INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2024
·Vol. 26
·期 1
·77
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IF 4.9
Zhiting Guo,Huiyan Yu,Kexin Yang,Wenjing Feng,Miao Liu,Tao Wang,Rong Xiao
Oxysterols, as metabolites of cholesterol, play a key role in cholesterol homeostasis, autophagosome formation, and regulation of immune responses. Disorders in oxysterol metabolism are closely related to the pathogenesis of neurodegenerative diseases. To systematically investigate the profound molecular regulatory mechanisms of neurodegenerative diseases, it is necessary to quantify oxysterols and their metabolites in central and peripheral biospecimens simultaneously and accurately. However, there are a lot of unsolved problems with the existing methods, such as the hindrance of applying a single method to different biological specimens or the challenge of simultaneous quantification due to differential groups on the ends of the oxysterol side chains. Herein, according to the physicochemical properties and structure of oxysterols, an optimized liquid chromatography-tandem mass spectrometry method for the quantification of oxysterols was established by optimizing the sample preparation process, chromatographic conditions, mobile phase pH, and solvent selection. Seven oxysterols were detected by this method, including 27-hydroxycholesterol, 7α-hydroxycholesterol, 7α,27-dihydroxycholesterol, 7-dehydrocholesterol, 7α-hydroxy-3-oxo-4-cholestenoic acid, 3-hydroxy-5-cholestenoic acid, and 24(S)-hydroxycholesterol. Non-derivatization extraction with methyl tert-butyl ether was used for different biospecimens, followed by simultaneous chromatographic separation of oxysterols on a phenyl hexyl column. By repeated validation, this method exhibited satisfactory linearity, precision, recovery, sensitivity, repeatability, and stability, and it was successfully applied to the detection of oxysterols in the plasma, cerebral cortex, and liver of mouse. In summary, our optimized method enables concurrent analysis and quantification of oxysterols and their metabolites in various biospecimens, presenting a broad range of applicability.
研究领域:
生物化学
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医学分析
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神经科学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Immunology
·
2024
·
IF 5.7
Li Na,Wang Tianyi,Zhang Huiying,Li Xiao,Bai Haochen,Lu Ning,Lu Kaizhi
Objective: This investigation sought to delineate the causal nexus between plasma glutamine concentrations and leukemia susceptibility utilizing bidirectional Mendelian Randomization (MR) analysis and to elucidate the metabolic ramifications of asparaginase therapy on glutamine dynamics in leukemia patients.Methods: A bidirectional two-sample MR framework was implemented, leveraging genetic variants as instrumental variables from extensive genome-wide association studies (GWAS) tailored to populations of European descent. Glutamine quantification was executed through a rigorously validated Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS) protocol. Comparative analyses of glutamine levels were conducted across leukemia patients versus healthy controls, pre- and post-asparaginase administration. Statistical evaluations employed inverse variance weighted (IVW) models, MR-Egger regression, and sensitivity tests addressing pleiotropy and heterogeneity.Results: The MR findings underscored a significant inverse association between glutamine levels and leukemia risk (IVW p = 0.03558833), positing lower glutamine levels as a contributory factor to heightened leukemia susceptibility. Conversely, the analysis disclosed no substantive causal impact of leukemia on glutamine modulation (IVW p = 0.9694758). Notably, post-asparaginase treatment, a marked decrement in plasma glutamine concentrations was observed in patients (p = 0.0068), underlining the profound metabolic influence of the therapeutic regimen.Conclusion: This study corroborates the hypothesized inverse relationship between plasma glutamine levels and leukemia risk, enhancing our understanding of glutamine’s role in leukemia pathophysiology. The pronounced reduction in glutamine levels following asparaginase intervention highlights the critical need for meticulous metabolic monitoring to refine therapeutic efficacy and optimize patient management in clinical oncology. These insights pave the way for more tailored and efficacious treatment modalities in the realm of personalized medicine.
研究领域:
医学
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遗传学
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代谢组学
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产品关联
DOI
PubMed
高影响因子
CLINICA CHIMICA ACTA
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2021
·
IF 3.786
Qian Gong,Xiuming Zhang,Aifeng Liang,Sinian Huang,Guangang Tian,Mengjiao Yuan,Qing Ke,Yijun Cai,Bin Yan,Jin Wang,Jinjin Wang
Background and aims Colorectal cancer (CRC) is part of the most widespread malignant tumors. At present, colonoscopy is a routine procedure in the diagnosis of CRC, but it is traumatic. Carcinoembryonic antigen, CA199, and CA242 are common serum markers for the diagnosis of CRC; however, they do not demonstrate satisfactory specificity and sensitivity for the diagnosis of CRC. Hence, Now it is necessary to screen many valuable serum biomarkers for CRC, proteomics methods have been used to investigate PTMs such as glycosylation of proteins with prominent roles in the occurrence and development of tumors. Methods This study screens altering glycosylated proteins of CRC tissues using LC-MS/MS quantitative glycoproteomics, and then these candidate biomarkers for CRC are further validated by serum glycoproteomics. Results The results of glycoproteomics in CRC tissues show that the abundance of 160 and 79 glycerogelatin proteins was obviously upregulated and downregulated compared with their adjacent tissues( P < 0.05). Bioinformatics analysis suggests that these molecules are mainly involved in many biological processes, including skeletal system development, collagen fibril organization, and receptor-mediated endocytosis. Results of serum glycoproteomics show that the changing trends of 2 protein glycosylation were consistent with MS results of CRC tissues, including ICAM1and APMAP. Areas under the ROC curve (AUC) results confirm that ICAM1and APMAP as early immune diagnosis markers of CRC has excellent sensitivity and specificity. Conclusion The ICAM1 and APMAP may serve as a potential tumor marker for CRC.
研究领域:
肿瘤学
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蛋白质组学
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医学
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DOI
PubMed
高影响因子
International Journal of Analytical Chemistry
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2020
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IF 1.678
Fang Zhao,Yuan Wei,Yiming Yan,Han Liu,Sitong Zhou,Bo Ren,Ruijuan Liu
Toxicity induced by crizotinib, a small-molecule tyrosine kinase inhibitor, is a significant clinical issue during treatment. A tissue distribution study is required to explore the organs affected by this molecule. In this study, a simple liquid chromatography tandem mass spectrometry method was developed and validated for the determination of crizotinib in various mouse tissues. Mouse tissue homogenates were processed by protein precipitation with methanol, and apatinib was chosen as the internal standard. The analytes were separated on a Phenomenex Kinetex C18 (50 mm × 2.1 mm, 2.6 μm) column with gradient elution using methanol and 0.3% formic acid water solution. Tandem mass spectrometric detection was conducted using multiple reaction monitoring via an electrospray ionization source in the positive mode. The monitored ion transitions were m/z 450.1 ⟶ 260.2 for crizotinib and m/z 398.2 ⟶ 212.0 for apatinib. The problem of the severe carryover effect was successfully resolved. The method was validated and applied to a tissue distribution study of crizotinib in mice, which was reported for the first time. The results of the study showed that the main target organs of crizotinib were the lung, liver, and spleen, and a high concentration of crizotinib was found in the gastrointestinal tract. This study offers a reliable method for quantifying crizotinib and provides a basis for further research on crizotinib toxicity.
研究领域:
药理学
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分析化学
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毒理学
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DOI
PubMed
高影响因子
Food Analytical Methods
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2016
·Vol. 10
·期 5
·1386-1396
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IF 2.167
Wang Yu,Gao Lei,Qin Dongli,Chen Ligang
In this work, a simple and selective method based on matrix solid phase dispersion (MSPD) using molecularly imprinted polymers (MIPs) as sorbents was successfully developed for the determination of melamine (MEL) in milk powder. The MIPs have been prepared by surface imprinted polymerization method using carbon nanotube (CNT) as support, MEL as template molecule, γ-aminopropyltriethoxysilane as functional monomer, and tetraethyl orthosilicate as cross-linking agent. The MIPs were characterized by Fourier transform infrared spectrometer and scanning electron microscopy. At the same time, the adsorption isotherm, dynamic adsorption, and selective adsorption of MIPs to MEL were also studied. The prepared MIPs were used as dispersant of MSPD to extraction and separate MEL in milk powder. We optimized the extraction conditions as follows: the ratio of MIPs to sample was 2:3; the dispersion time was 10 min; washing solvent was 2.0 mL acetonitrile; and elution solvent was 4.0 mL ammonium methanol (5:95, v / v ). Once the MSPD process was completed, the MEL extracted from milk powder was determined by LC-MS/MS. Under the optimal conditions, the limit of detection of MEL was 0.2 ng g −1 . The relative standard deviations of intra-day and inter-day were obtained in the range of 0.4∼5.1 %. The satisfied recoveries of MEL were 92.5∼97.2 %. The MIP-MSPD method has some advantages, such as higher selectivity, lower cost of preparation, less time consuming and organic solvent consumption, simpler sample pretreatment procedure, and higher extraction recovery compared with those existing methods which were used for determination of MEL.
研究领域:
食品科学
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分析化学
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产品关联
DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
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2017
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IF 2.603
Lingli He,Chuan Li,Xuyuan Liu,Qingqing Yang,Haizhi Zhang,Weiren Xu,Luyong Zhang,Changxiao Liu
Benzbromarone is a uricosuric drug metabolized predominantly by cytochrome P450 2C9 from in vitro findings. Human CYP2C9 exhibits extensive genetic polymorphism and numbers of clinic studies have demonstrated that CYP2C9 genetic polymorphism has a significant influence on the pharmacokinetics of benzbromarone. But in vitro study on the interaction between CYP2C9 allelic isoforms and benzbromarone was rare. Here, an LC–MS/MS method was established and validated to determine the concentration of benzbromarone in different CYP2C9 enzyme incubation systems for the drug-enzyme interaction study. By selecting appropriate internal standard and optimizing separation system, including mobile phase , sample solvent and gradient elution condition, this LC–MS/MS method was developed with fine linearity (r 2 ≥ 0.996), good reproducibility (RSD ≤ 6.6%), high stability (92.37–114.67%), efficient recovery (91.23–109.82%) and acceptable matrix effect (110.54–115.31%). Based on this method, the interaction between 3 CYP2C9 allelic isoforms and benzbromarone was researched by kinetics parameters (K m , V max , Cl int ). As a result, CYP2C9*1 displayed the highest metabolic activity towards benzbromarone, CYP2C9*2 showed a little lower catalytic activity than CYP2C9*1 (relative clearance/*1 = 85.86%), CYP2C9*3 showed the lowest catalytic activity (relative clearance/*1 = 21.57%). The result illustrated that various CYP2C9 allelic isoforms showed different enzymatic activities towards benzbromarone, which could offer effective consultation for personalized administration in clinic.
研究领域:
药理学
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医学遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
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2021
·
IF 7.514
Libin Wan,Haidong Gao,Huoliang Gao,Rui Du,Fayun Wang,Yong Wang,Mantang Chen
For the first time, electromembrane extraction (EME) combined LC-MS/MS was applied to extract and determine α-solanine and α-chaconine in different potato tissues using NPOE containing 20% (v/v) DEHP as supported liquid membrane (SLM). Under the optimal conditions, the proposed EME-LC-MS/MS method was evaluated using spiked fresh potato peel sample. The linear range for α-solanine and α-chaconine was 5–1000 ng mL −1 (R 2 > 0.9991), with LOD and LOQ of 1.2–1.5 ng mL −1 and 4.1–5.2 ng mL −1 , respectively. Repeatability for α-solanine and α-chaconine at three concentration levels was satisfactory (<4.9%), and recoveries ranged from 73% to 106%. Finally, the EME-LC-MS/MS method has been successfully employed to determine α-solanine and α-chaconine in sprouted potato peel and tuber samples, indicating that EME exhibited high selectivity and efficient sample clean-up capability. Consequently, EME showed great potential for extraction and purification of toxic and bioactive basic compounds from complex plant tissues.
研究领域:
分析化学
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植物化学
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食品安全
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产品关联
DOI
PubMed
高影响因子
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
·
2022
·Vol. 37
·期 1
·e9397
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IF 2.586
Wenrui Yao,Jinfeng Ge,Qiaozhuan Hu,Jingying Ma,Daohe Yuan,Xiaoli Fu,Yulin Qi,Dietrich A. Volmer
Rationale The development of appropriate analytical screening techniques for pharmaceuticals and personal care products (PPCPs) is the basis for studying the distribution and environmental impact of emerging contaminants (ECs). Mass spectrometry-based screening methods vary with the complexity of the target compounds. It is challenging to balance both positive and negative ion quantification with a low detection limit. To establish a set of experimental methods including extraction, chromatography-separation and mass spectrometry screening is one of the most important topics in PPCP research. This paper describes a universal and efficient qualification and quantification protocol for the simultaneous detection of 34 PPCPs in different environmental samples in a single analytical data acquisition run. Methods Thirty-four representative PPCPs, which are widely distributed in the environment with high ecological toxicity and complex chemical structures, were selected as representative target ECs. The extraction of the target PPCPs was achieved using only one solid-phase extraction cartridge without the need to adjust the pH of samples. The enriched samples were detected by LC–MS/MS in both positive and negative ion modes simultaneously. The protocol was evaluated based on the accuracy, precision, detection limits and matrix effects. Results This method achieved simultaneous detection of PPCPs in both positive and negative ion modes, with a single analytical cycle of 12 min. The observed SPE recoveries were between 40% and 115%. The instrumental detection limits (IDL) varied from 0.01 to 1 pg, and the method detection limits (MDL) were between 0.002 and 3.323 ng/l in different matrices. Most of the PPCPs were subjected to matrix suppression below 30%. The method was successfully applied for quantitative analysis of the PPCPs in different environmental samples, including river samples, wastewater treatment plant (WWTP) samples and soil samples. Conclusions This protocol developed a rapid and efficient detection method to simultaneous qualitative and quantitative 34 representative PPCPs in the environment. The IDL ranged from 0.01 to 1 pg and the MDL ranged from 0.002 to 3.323 ng/l in different matrices. The detection limit was one order of magnitude lower compared to previous studies. The protocol also provided a wide application range for different environmental matrices, which permitted the migration and transformation of PPCPs to be explored.
研究领域:
环境科学
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分析化学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Oncology
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2025
·
IF 3.3
Yu-jiao Hua,Ying Zhang,Rui-Rong Wu,Juan Lv,Yan Zhang,Yan-yan Chen,Yong-juan Ding,Jing-hua Chen
Background Oxaliplatin-induced peripheral neuropathy (OIPN) poses a significant challenge for patients with colorectal tumor, often resulting in treatment interruption or discontinuation and subsequent treatment failure. Herein, a longitudinal untargeted metabolomic study to reveal the metabolomic profiles and biomarkers associated with the progression of OIPN. Methods A prospective cohort of 129 colorectal cancer patients receiving oxaliplatin-based chemotherapy was stratified into four OIPN severity grades (Level 0-3). Serum samples underwent untargeted LC-MS/MS metabolomic analysis, detecting 521 metabolites. Multivariate statistical models and SHAP-guided random forest algorithms were employed to prioritize biomarkers. Machine learning validation included six classifiers assessed via ROC-AUC. Results The cumulative dose of Oxaliplatin chemotherapy plays an important role in OIPN. At the same time, our findings implied that the occurrence of OIPN may be associated with the progression of the disease and the patients’ tumor markers (CEA, CA19-9, CA72-4), as well as immune response and inflammation (ANC, PLT), and metabolic and liver function abnormalities (GGT and UA) (P<0.05).Multivariate statistical analysis combined with SHAP-guided machine learning identified six biomarkers, including thiabendazole, 1-methylxanthine, imidazol-5-yl-pyruvate, 5-hydroxypentanoic acid, spermidine, and 4’-oxolividamine that consistently distinguished OIPN patients (Level 1-3) from non-OIPN controls (Level 0). Machine learning models, validated across six classifiers, demonstrated near-perfect discrimination for early-stage OIPN (AUC nearly 1). However, differentiation between intermediate OIPN grades (Level 1 vs 2, Level 1 vs 3, Level 2 vs 3) yielded lower predictive accuracy (AUC: 0.549–0.843), likely due to cohort size limitations and reliance on subjective sensory-based grading. Pathway enrichment analysis highlighted dysregulation in ABC transporters, central carbon metabolism in cancer, amino acid metabolism, and linoleic acid metabolism, suggesting potential roles in OIPN pathogenesis. Conclusions These findings suggest that the selected biomarkers could serve as a foundation for the prediction and management of OIPN in colorectal cancer patients.
研究领域:
肿瘤学
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神经毒理学
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转化医学
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药物代谢
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代谢组学
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精准医学
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临床药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CLINICA CHIMICA ACTA
·
2026
·
IF 4.1
Zhenni Liu,Xiaohua Xiao,Bingchu Li,Nijiao Qin,Ziyang Li,Xing Lyu,Zhongyuan Xiang,Min Hu,Qichen Long
A rapid and practical LC-MS/MS method for ascitic-fluid CK18 quantification. • Sequential salting-out/pI-guided acidic precipitation strategy for CK18 enrichment. • Simplified pretreatment shortens overall sample preparation duration to merely 6 h. • Ascitic-fluid CK18 correlates with hepatic markers for auxiliary cirrhosis evaluation. Cytokeratin 18 (CK18), a predominant hepatocyte intermediate filament protein released during hepatocyte death, has been investigated as a hepatic injury biomarker. Conventional immunoassays for CK18 suffer from non-specific cross-reactivity and insufficient standardization. Though liquid chromatography-tandem mass spectrometry (LC-MS/MS) could overcome such immunoanalytical drawbacks, accurate CK18 quantification in ascitic fluid remains analytically challenging because of the complex matrix, low target abundance, and variability introduced by protein enrichment and digestion. Herein, we developed a rapid and practical LC-MS/MS method for CK18 quantification in ascitic fluid. A two-step non-immunoaffinity enrichment strategy, integrating sequential ammonium sulfate salting-out and isoelectric point-driven acidic precipitation, was optimized to balance enrichment yield, pretreatment turnaround, and operational cost. Subsequent pretreatment workflow comprising urea denaturation, enzymatic hydrolysis, and solid phase extraction purification can be completed within 6 h, providing an economical, instrument-compatible procedure for LC-MS/MS analysis. Systematic analytical performance evaluation showed satisfactory linearity spanning 0.10–10.36 nmol/L, a limit of quantification of 0.10 nmol/L, a limit of detection of 0.02 nmol/L, total imprecision <10%, and recoveries ranging from 93.07% to 108.92%. The internal standard effectively compensated for LC-MS/MS instrumental variability and ionization-related matrix effects. Method comparison showed acceptable agreement with a published multiplexed cytokeratin MS method. Clinical analysis demonstrated significantly elevated ascitic-fluid CK18 concentration in cirrhotic patients versus non-cirrhotic controls ( P = 0.001). This novel enrichment strategy preserves the simplicity and cost-efficiency of conventional precipitation methods while avoiding costly antibody capture reagents. Collectively, the validated LC-MS/MS method is appropriate for preliminary exploration of the clinical value of ascitic-fluid CK18 and can facilitate translational research in cirrhotic populations.
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DOI
高影响因子
Drug Design Development and Therapy
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2026
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IF 6.1
Na Yang,Yuqing Zhao,Shiyu Liu,Yifan Zhou,Xinyue Ji,Tianqi Zhang,Runbin Sun,Huaijun Zhu,Yao Du,Min Wang
Background Mycophenolic acid (MPA), administered as mycophenolate mofetil (MMF) or enteric-coated mycophenolate sodium (EC-MPS), is the first-line immunosuppressant for kidney transplant patients. Traditional plasma-based therapeutic drug monitoring (TDM) for MPA fails to accurately reflect intracellular drug exposure at the pharmacological action site.Purpose This study aimed to establish and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous quantification of MPA and its glucuronide metabolite, MPAG, in peripheral blood mononuclear cells (PBMCs) to support cellular pharmacokinetic (PK) assessment.Methods Chromatographic separation was performed on a BEH C18 column using methanol-water containing 0.1% formic acid and 5 mmol/L ammonium acetate. A total of 139 PBMC samples from 40 kidney transplant recipients (n=19 receiving EC-MPS; n=21 receiving MMF) were analyzed to evaluate the plasma-PBMC PK correlations and gender-related differences.Results The method exhibited excellent linearity over the concentration range of 0.2–500 ng/mL (R2 > 0.998), along with acceptable precision, accuracy and matrix effect. The analytes remained stable under various experimental conditions. Significant inter-individual variability in intracellular MPA and MPAG concentrations was observed among patients. Spearman correlation analysis demonstrated a weak-to-moderate positive correlation between plasma and intracellular PBMC concentrations of MPA (EC-MPS, Rs=0.396; MMF, Rs=0.480). Additionally, the concentration-time profiles of MPA and MPAG in PBMCs of kidney transplant recipients after administration of EC-MPS or MMF exhibited different characteristics. No significant gender-based differences in plasma and PBMCs were observed.Conclusion We established and validated an LC-MS/MS method for simultaneous quantification of intracellular MPA and MPAG. The observed weak-to-moderate correlation between plasma and PBMC exposure highlights the inherent limitations of conventional plasma-based TDM for MPA. Collectively, our study provides a feasible approach for monitoring intracellular drug levels and facilitates the optimization of individualized immunosuppressive dosing regimens for renal transplant recipients.
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DOI
高影响因子
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
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2026
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IF 2.5
Yagang Li,Shuyan Huo,Yunhui Xing,Yonghang He,Wei Wu,Min Huang,Yiming Jiang,Guoping Zhong,Weiwei Zeng
Novel TiO 2 @MWCNTs composites were designed and synthesized, which exhibit potential as purification materials for phospholipid removal from rat plasma samples. • A dispersive solid-phase extraction method based on TiO 2 @MWCNTs showed the capacity to mitigate matrix effects in rat plasma samples for small-molecule analytes with low lg P . • An LC–MS/MS method was established and validated for quantifying representative drugs using TiO 2 @MWCNTs and applied to a pharmacokinetic study of metformin. The matrix effect is a prevalent interference in LC–MS/MS bioanalysis of small-molecule drugs, with lipids as a major source of such interference in plasma matrices. Leveraging the lipid-adsorbing capability of multi-walled carbon nanotubes (MWCNTs), this study synthesized TiO 2 @MWCNTs composites and developed a novel dispersive solid-phase extraction (d-SPE) method to reduce matrix effects in rat plasma. Lipid profiles were compared to characterize the lipid purification capacity of TiO 2 @MWCNTs in rat plasma samples. The matrix effects of representative small-molecule drugs, including metformin, amisulpride, and aripiprazole, were compared across TiO 2 @MWCNTs-based d-SPE, protein precipitation, and traditional solid-phase extraction. Method validation and sample determination were subsequently performed. The results demonstrated that TiO 2 could be successfully loaded onto MWCNTs. The responses of 72 differential lipid ions were significantly reduced by at least 2-fold after TiO 2 @MWCNTs treatment, and the IS-normalized matrix factors for metformin (from 13.69 to 20.00% to 31.49–44.39%) and amisulpride (from 71.01 to 84.82% to 91.69–101.48%) as well as accuracy for aripiprazole at low concentration were significantly improved compared with protein precipitation treatment. Method validation results confirmed excellent selectivity, accuracy, precision, and reproducibility. The validated method was also successfully applied to a pharmacokinetic study of metformin with pharmacokinetic parameters comparable to those reported previously. This study shows that the dispersive solid-phase extraction method based on TiO 2 @MWCNTs could act as a simple, rapid, low-cost and reliable pretreatment strategy to mitigate matrix effects for small-molecule analytes with low lg P in rat plasma as a proof-of-concept attempt.
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DOI
高影响因子
危险品化学品经营许可证（带存储）