INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 26
·期 1
·77
·
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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JOURNAL OF CHROMATOGRAPHY A
·
2026
·
IF 4
Chenyue Zhu,Chuanxiang Yao,Huiting Qin,Wei Zhang,Xinyue Zheng,Siying Zheng,Xinyuan Ye,Ying Xiong,Lili Jiang,Sen Li,Meiqin Wang,Hongliang Jiang,Zhifeng Du
The presence of deamination impurities in phosphorothioate oligonucleotides (PS-ONs) presents a significant challenge to quality control, as their detection is complicated by minimal mass differences from the parent molecule and difficulties in chromatographic separation. Herein, a bottom-up strategy based on enzymatic digestion, chemical derivatization, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was proposed for the sensitive qualitative and quantitative analysis of trace cytosine deamination in PS-ONs. Fomivirsen (FMVS) and its deamination impurity were used as models. The method involved digestion with S1 nuclease, followed by derivatization of the 5’-phosphorothioate in digests with N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTBSTFA) to enhance chromatographic retention and mass spectrometric response. This approach effectively eliminated the need for ion-pairing (IP) reagents. The mass shift upon deamination was amplified in the smaller digested fragments, and a unique isotopic pattern in the derivatized products, specifically, the decrease of the first isotopic peak and the increase of the second isotopic peak was exploited for quantification. The method demonstrated excellent linearity over a weight ratio of deamination product to prototype range from 0.05:1 to 0.9:1. Satisfactory accuracy (80–120%) and precision (RSD < 15%) were achieved. Furthermore, the precise deamination site (C8) within a sample containing 10% deamination impurities was successfully identified. Thus, this method provides a powerful tool for profiling deamination impurities in PS-ONs, with potential applications in quality control, stability studies, and the characterization of other nucleic acid modifications.
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EUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY
·
2026
·Vol. 128
·期 9
·e70142
·
IF 2.4
Yunping Yao,Zehui Zhang,Bingxin Li,Changmo Li
Accurate qualitative and quantitative determination of free radicals during lipid oxidation, which is the core process of edible oil quality deterioration and essentially a free radical chain reaction, is of crucial importance. To overcome this problem, we developed and validated an analytical method for the specific characterization of key radicals in lipid oxidation. These radicals (R•, RO•, and ROO•) were trapped using N-tert -butyl-α-phenylnitrone under iron-mediated conditions, leading to the formation of stable adducts that were subsequently analyzed by LC–MS/MS–IT–TOF. This method offers good sensitivity with low limits of detection (0.06–0.15 µM) and limits of quantification (0.20–0.50 µM). Density functional theory calculations further confirm that the reaction proceeds readily and yields thermodynamically stable products. When applied to the methyl linoleate oxidation system, the method demonstrated that methyl linoleate alkyl radical (L•) predominated during oxidation, with its concentration peaking at 3.74 × 10 −2 mM (accounting for up to 70% of radicals) at 4 h followed by a decline due to chain termination reactions, whereas methyl linoleate peroxyl radicals (LOO•) concentration stabilized as oxidation progressed (1.46 × 10 −2 mM, 27% of radicals), and methyl linoleate alkoxyl radicals (LO•) remained at a consistently low concentration (1.58 × 10 −3 mM) owing to its preferential conversion via the β-scission pathway. These results were consistent with electron spin resonance (ESR) data, thereby validating the method's reliability. Consequently, this method enables the precise monitoring of lipid oxidation processes, serving as a valuable tool for assessing antioxidant efficacy in lipid science. Practical Applications : We present an analytical method integrating N-tert -butyl-α-phenylnitrone (PBN)-based spin trapping with LC–MS/MS–IT–TOF for direct tracking of key lipid radicals (L•, LO•, and LOO•) throughout the lipid oxidation process. This method addresses a long-standing technical gap in lipid oxidation research, providing a robust, practical tool for both high-throughput screening of high-efficacy antioxidants and rational design of stable lipid-containing food formulations. By bridging fundamental analysis and product development, our technique has clear real-world implications for extending the shelf life of lipid-based foods, offering significant value to food chemistry research while delivering substantial economic and sustainability benefits to the global food industry.
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SAUDI PHARMACEUTICAL JOURNAL
·
2025
·Vol. 33
·期 5
·1-17
·
IF 3.4
Wang Ziheng,Xu Yuchen,Liu Manting,Yang Yuchang,Shi Wenjuan,Zhu Qian,Liu Juan,Yi Lisha,Wu Huimin,Yin Xingbin,Dong Xiaoxv,Ni Jian,Qu Changhai
This study aimed to establish a method for quantifying baicalin (BC) in rabbit ocular tissues and plasma, and evaluate the pharmacological efficacy and pharmacokinetic properties of BC and BC@HS15/DSPE-PEG2000-L-Val, a novel ocular formulation for dry eye treatment. BC@HS15/DSPE-PEG2000-L-Val or free BC was administered via eye drops to benzalkonium chloride (BAC)-induced dry eye mice. Corneal and conjunctival tissues were assessed for anti-dry eye efficacy. BC concentrations in cornea, conjunctiva, aqueous humor, and ocular plasma were quantified using LC–MS/MS. Noncompartmental pharmacokinetic parameters (AUC, Tmax) were calculated using DAS 2.0 software. The method demonstrated excellent linearity (0.50–500.00 ng/mL, r > 0.9905), precision (RSD < 10%), and accuracy (± 13%). Compared to free BC, BC@HS15/DSPE-PEG2000-L-Val significantly increased tear secretion, reduced MMP-3/MMP-9 expression, and preserved corneal epithelium integrity. In the micelle group, corneal and conjunctival Cmax values were 2.7- and 3.6-fold higher than the solution group, respectively. A sensitive and validated liquid chromatography-tandem mass spectrometry (LC–MS/MS) method was developed to measure baicalin concentrations in ocular plasma and tissues of rabbits. BC@HS15/DSPE-PEG2000-L-Val for treating dry eye demonstrated significantly superior outcomes. The nano-micelle notably enhanced BC concentration on the ocular surface and effectively prolonged its retention time. Graphical abstract
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Food Analytical Methods
·
2016
·Vol. 10
·期 5
·1386-1396
·
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
·
分析化学
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JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
·
2017
·
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
·
医学遗传学
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RAPID COMMUNICATIONS IN MASS SPECTROMETRY
·
2022
·Vol. 37
·期 1
·e9397
·
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.
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FOOD CHEMISTRY
·
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.
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International Journal of Analytical Chemistry
·
2020
·
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.
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LWT-FOOD SCIENCE AND TECHNOLOGY
·
2024
·
IF 6
Li Liang,Jincheng Zhang,Baoguo Sun,Ming Zhang,Yan Zhang,Ku Li,Yuyu Zhang
Hypertension has emerged as a major factor detrimental to human health. Umami peptide IIVFGRQLL from yeast has a certain ACE inhibitory activity (IC 50 : 59.35 μM) at 0.14 mg/mL. Moreover, IIVFGRQLL could regulate the systolic blood pressure of spontaneously hypertensive rats model indicating its potential antihypertensive effect. The changes in ACE inhibitory activity and peptide sequence during the digestion were investigated based on the ACE inhibition assays, simulated gastrointestinal digestion, and LC-MS/MS. The ACE inhibitory activity of IIVFGRQLL decreased by 39.0% after digestion. It was hydrolyzed into 5 peptide fragments, among which IVF took the biggest share followed by IVFG, VF, FGR, and RQL. The inhibitory rate of IVF (78.70%), FGR (86.30%), and VF (83.50%) was higher than that of IIVFGRQLL (60.11%). The molecular docking results revealed all the peptides interacted with active residues related to Zn 2+ and S1 pocket in ACE. Besides, IVF, FGR and RQL showed increased electrostatic interaction with Zn 2+ . These results provide theoretical support for the future exploration of umami peptide as a food additive or seasoning to alleviate hypertension.
研究领域:
药理学
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食品科学
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营养学
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生物化学
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Scientific Reports
·
2015
·Vol. 5
·期 1
·1-11
·
IF 5.578
Yang Ting,Xu Feifei,Fang Danjun,Chen Yun
The distinct roles of protein isoforms in cancer are becoming increasingly evident. FRα and FRβ, two major isoforms of the folate receptor family, generally have different cellular distribution and tissue specificity. However, the presence of FRβ in breast tumors, where FRα is normally expressed, complicates this situation. Prior to applying any FR isoform-based diagnosis and therapeutics, it is essential to monitor the expression profile of FR isoforms in a more accurate manner. An LC-MS/MS-based targeted proteomics assay was developed and validated in this study because of the lack of suitable methodology for the simultaneous and specific measurement of highly homologous isoforms occurring at low concentrations. FRα and FRβ monitoring was achieved by measuring their surrogate isoform-specific peptides. Five human breast cell lines, isolated macrophages and 60 matched pairs of breast tissue samples were subjected to the analysis. The results indicated that FRβ was overexpressed in tumor-associated macrophages (TAMs) but not epithelial cells, in addition to an enhanced level of FRα in breast cancer cells and tissue samples. Moreover, the levels of the FR isoforms were evaluated according to the histology, histopathological features and molecular subtypes of breast cancer. Several positive associations with PR/ER and HER2 status and metastasis were revealed.
研究领域:
癌症生物学
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蛋白质组学
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临床肿瘤学
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Scientific Reports
·
2025
·Vol. 15
·期 1
·1-8
·
IF 3.9
Liu Hao,Cai Meng,Peng Yong,Miao Jing-kun,Yu Jin
While remifentanil, etomidate, and rocuronium are increasingly used for cesarean section due to their favorable hemodynamic stability and fetal safety profile, their pharmacokinetics and potential effects on neonates remain poorly understood. This study developed and validated a rapid, sensitive LC–MS/MS method for simultaneous quantification of the three anesthetics in microsamples of 10 μl serum, followed by a paired maternal and umbilical cord serum investigation. After protein precipitation with acetonitrile, analytes were separated within 4 min using positive electrospray ionization in MRM mode. Method validation demonstrated excellent linearity (R 2 > 0.99) for all compounds, with LLOQs of 0.15 ± 0.02 ng/mL (remifentanil), 16.87 ± 0.51 ng/mL (etomidate), and 106.73 ± 8.63 ng/mL (rocuronium). Precision (intra-/inter-day < 15%) and minimal carry-over (< 5%) met bioanalytical standards. Applied to 20 maternal-newborn pairs, the method quantified differential drug distribution: maternal arterial concentrations (remifentanil 4.75 ± 0.19 ng/mL; etomidate 412.71 ± 35.29 ng/mL; rocuronium 7.08 ± 0.48 μg/mL) exceeded umbilical vein levels (2.43 ± 0.13 ng/mL; 302.15 ± 29.03 ng/mL; 0.86 ± 0.16 μg/mL), which were higher than umbilical artery concentrations (1.33 ± 0.15 ng/mL; 166.24 ± 21.53 ng/mL; 0.44 ± 0.77 μg/mL). Calculated placental transfer rates significantly differed among anesthetics (remifentanil 0.52 ± 0.02; etomidate 0.75 ± 0.04; rocuronium 0.13 ± 0.02; all P < 0.001), reflecting distinct pharmacokinetic behaviors. The validated method enables reliable microvolume analysis for perinatal pharmacokinetic studies, particularly valuable when sample availability is limited. Its rapid throughput and sensitivity make it suitable for clinical research applications investigating maternal–fetal drug transfer dynamics.
研究领域:
分析化学
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药代动力学
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麻醉学
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妇产科学
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新生儿学
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临床研究
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