MOLECULES
·
2023
·Vol. 28
·期 5
·2415
·
IF 4.6
Ao Xu,Yunlin Xue,Yuyu Zeng,Jing Li,Huiling Zhou,Zhen Wang,Yin Chen,Hui Chen,Jian Jin,Tao Zhuang
Furosemide is a widely used loop diuretic in the treatment of congestive heart failure and edema. During the preparation of furosemide, a new process-related impurity G in the levels ranging from 0.08% to 0.13% was detected in pilot batches by a new high performance liquid chromatography (HPLC) method. The new impurity was isolated and characterized by comprehensive analysis of FT-IR, Q-TOF/LC-MS, 1D-NMR (1H,13C, and DEPT), and 2D-NMR (1H-1H-COSY, HSQC, and HMBC) spectroscopy data. The possible formation pathway of impurity G was also discussed in detail. Moreover, a novel HPLC method was developed and validated for the determination of impurity G and the other six known impurities registered in the European Pharmacopoeia as per ICH guidelines. The HPLC method was validated with respect to system suitability, linearity, the limit of quantitation, the limit of detection, precision, accuracy, and robustness. The characterization of impurity G and the validation of its quantitative HPLC method were reported for the first time in this paper. Finally, the toxicological properties of impurity G were predicted by the in silico webserver ProTox-II.Keywords:furosemide;process-related impurity;characterization;Q-TOF/LC–MS and NMR;method validation
研究领域:
药物化学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
RAPID COMMUNICATIONS IN MASS SPECTROMETRY
·
2021
·Vol. 36
·期 8
·e9242
·
IF 2.419
Bin Xu,Pengfei Jia,Jun Cai,Lidong Gu,Huaxiao Yan,Hui Zhao,Song Qin
Rationale Saliva has been widely accepted as a more convenient alternative to serum or plasma in the field of clinical diagnosis. However, the detection of trace components in saliva has been a bottleneck problem. The aim of this work was to develop a highly sensitive and reliable method for simultaneously determining the trace steroid hormones including some with poor ionization efficiency in human saliva by liquid chromatography/tandem mass spectrometry (LC/MS). Methods Saliva was deproteinated by acetonitrile containing mixed isotope internal standards and extracted with methyl tert -butyl ether. The extraction solution was dried under a stream of nitrogen and the residue was derivatized using 50 mM O -ethylhydroxylamine hydrochloride in 80% methanol/water solution (v/v). The processed sample was determined by LC/MS in multiple reaction monitoring (MRM) mode. Results The method was successfully established for the simultaneous quantification of seven steroid hormones in human saliva and showed excellent specificity and sensitivity. The limits of quantification (LOQs) of all steroid hormones were below 5 pg/mL, in particular, the LOQ of progesterone was as low as 0.15 pg/mL. The linear correlation coefficients (r) were greater than 0.9990 in the range of 2–200 pg/mL for T, DHEA, A4, P4, P5, and 17OHP4 and in the range of 5–500 pg/mL for 17OHP5. The intra-day and inter-day variability ranged from 1.86% to 7.83% and 1.95% to 10.4%, respectively. The recovery of the method ranged from 86.9% to 111.1% for all steroid hormones using three spiked concentrations. Conclusions A novel LC/MS/MS method was developed for the simultaneous quantification of seven kinds of trace steroid hormones in human saliva. The results of the methodological study showed that the method exhibited excellent sensitivity and reliability for the evaluation of free steroid hormones in the human body. It is believed that this method could provide useful information of steroid hormone metabolism for auxiliary diagnosis of some endocrine disorders.
研究领域:
临床诊断
·
生物化学
·
内分泌学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2018
·
IF 2.831
Mengzhe Guo,Dantong Liu,Qian Sha,Houfa Geng,Jun Liang,Daoquan Tang
The epigenetic modification of DNA or RNA may have relationship with the development of diabetic nephropathy (DN). We designed to develop a liquid chromatography mass spectrometry (LC–MS) coupled with mobile phase additive sensitization method for the analysis of the dissociative epigenetic modified nucleosides in serum of patients from different periods of DN with the health comparison. Serum samples were collected from 43 healthy volunteers and 156 patients, which divided into four groups. A succinic acid enhanced LC–MS/MS strategy was developed for the quantitative analysis of seven nucleosides in serum samples. The signal intensity of seven nucleosides increased three to seven times, respectively. A series of statistical analysis were carried out to demonstrate the dynamic changes of modified nucleosides in four groups. This method showed good specific, accuracy, and stability. The results were calibrated and presented through the formation of m 6 A/C, I/C, 5-mdC/C, 5-mC/C, pseU/U to eliminate the individual and age differences. The m 6 A/C, I/C, and pseU/U were found having the dynamic changes in four groups ( p < 0.05). And these five modified nucleosides can also used as one parameter to distinguish these four groups by ROC and LDA analysis. Additionally, they were found to be connected with several clinical biochemical indexes through multiple linear regression analysis. Our LC–MS/MS method assay successfully quantifies the modified nucleosides in serum. And three modified nucleosides can act as the potential biomarkers to describe the development of DN and help to evaluate the diagnosis, treatment, and prognosis of DN in clinical.
研究领域:
医学
·
分子生物学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2022
·
IF 5.895
Xiaohui Feng,Na Liu,Youyou Yang,Shengnan Feng,Juan Wang,Qingshi Meng
Short-chain fatty acids (SCFAs) are major gut microbiota-derived metabolites, which can reshape the intestine and regulate gut immunity. The application of conventional GC methods has been hampered for quantifying low-concentrated SCFAs, such as in serum, saliva, and digesta of germ-free animals. Herein, we established a LC–MS method to quantify SCFAs after 5-(dimethylamino)-1-carbohydrazide-isoquinoline (DMAQ) derivatization. The DMAQ derivatization significantly enhanced the detection sensitivity and improved separation of SCFAs. 2-methylbutyric acid and 3-methylbutyric acid were separately quantitated. Moreover, the matrix effect was diminished using DMAQ-13C/15N-tagged SCFAs as internal standards. The established quantitation method was successfully applied in the analysis of plasma and cecum digesta collected from neonatal piglets, revealing that significant increases in biological SCFA contents in cecum digesta were closely related to the variation of gut microbial diversity. The established quantitation method is capable of sensitively and comprehensively quantifying SCFAs that may provide insights into underlying gut-microbiota functions.
研究领域:
分析化学
·
微生物学
·
代谢组学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2018
·
IF 2.831
Teng-Fei Yuan,Juan Le,Yan Cui,Rui Peng,Shao-Ting Wang,Yan Li
Background Comprehensive serum sex hormone profiling is essential for monitoring the occurrence and development of many related diseases. However, the current methods for multi-class sex hormone detection were always lack of Standard Reference Material (SRM) certification and suffered from large sample consumption. For improvement, we developed a new liquid chromatography-tandem mass spectrometry (LC–MS/MS) method focused on SRM certification and minimization of serum consumption for simultaneous quantification of seven mainstream serum sex hormones including estrogens (estrone E1, estradiol E2 and estriol E3), androgens (testosterone T, androstenedione AD, dehydroepiandrosterone DHEA) and progestogens (progesterone P). Methods To achieve one-batch analysis, a straightforward strategy was designed and carefully optimized. Schematically, serum was firstly spiked with isotope-labeled internal standards. Then, liquid-liquid extraction was performed with methyl tert-butyl ether. After drying under nitrogen, dansyl chloride was introduced for derivatization. Finally, the mixture was submitted to LC–MS/MS for quantification. Results The limit of quantification was 0.005 ng/mL for E1, E2 and E3, 0.01 ng/mL for T, P and AD, 0.25 ng/mL for DHEA. Inter- and intra-assay CVs were less than 11.8%. The selectivity was proved satisfactory by interference spiking tests. With systematical SRM validation, the mean bias of −5.4 to 4.7% was observed, which indicated excellent method reliability. We found significant positive bias in chemiluminescence immunoassay (CLIA) detection comparing with current method, which promoted us to reconsider our previous results on sex hormone regulation in male patients with coronary atherosclerotic disease. After redetecting the related samples, modified and improved conclusions were proposed. Conclusions A LC–MS/MS method for multi-class serum sex hormone profiling was developed with SRM certification and minimized serum consumption. Taking advantages of such reliable method, the previous CLIA-based research findings on sex hormone regulation in male patients with coronary atherosclerosis were modified and improved after redetecting the same sample-pool.
研究领域:
医学检验
·
内分泌学
·
临床化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHROMATOGRAPHIA
·
2017
·Vol. 80
·期 12
·1733-1739
·
IF 1.402
Zhang Chaozheng,Zheng Yu,Ma Shenxi,Wu Zhiguo
Abstract Bile acids are essential materials in the fatty acid cycle. Thus, understanding the metabolism of bile acids is significant. In this study, LC–MS was employed to analyze the composition and content of bile acids in rat cecal contents. Six individual bile acids were determined simultaneously. The mobile phase consisted of acetonitrile and 10 mmol L −1 of ammonium acetate solution. The flow rate was 0.2 mL min −1 , and the total analysis time was 34 min. Under the developed conditions, all calibration curves for the six individual bile acids were achieved, and all correlation coefficients exceeded 0.99. Meanwhile, satisfactory results were obtained in the range of 50–500 µg g −1 for the six individual bile acids in the rat cecal contents. The LOD and LOQ of the bile acids were as low as 2.5 and 10 µg g −1 in the rat cecal contents, respectively, and the relative standard deviations of the peak area were within 1.25–2.01%. The dried residues after extraction were selected as the blank matrix. The overall recovery efficiencies of the bile acids in biological samples were increased to 97.2–103.2%. Therefore, the developed method can be used to quantify individual bile acids at extremely low levels in rat cecal contents. Graphical Abstract
研究领域:
生物化学
·
分析化学
✓ 已验证
DOI
高影响因子
BIOMEDICAL CHROMATOGRAPHY
·
2025
·Vol. 39
·期 3
·e6090
·
IF 1.7
Fang Wang,Wenfei Sun,Zheheng Ma,Furui Chu
Proteolysis targeting chimera (PROTAC) has been developed and currently enjoys widespread interest among the field of pharmaceutical manufacturing in recent decades. NX-5948 is an orally active PROTAC Bruton tyrosine kinase degrader, which shows anti-inflammatory and antitumor activities. In this study, a simple and fast bioanalytical method for the quantification of NX-5948 in beagle dog plasma was developed using liquid chromatography–tandem mass spectrometry (LC-MS/MS). A full method validation was performed according to regulatory guidelines. For the quantification, [M + H] + was formed using an electrospray ionization (ESI) source in the positive ion mode, and selective reaction monitoring (SRM) was employed using a triple quadrupole mass spectrometry. The monitored precursor-to-product transitions were m/z 807.5 > 790.5 for NX-5948 and m/z 812.4 > 452.1 for internal standard. A linear response was obtained at the concentration range of 0.5–200 ng/mL, with correlation coefficient > 0.99. The interday accuracy ranged from −9.03% to 5.99% (RSD < 7.84%), and the intraday accuracy from −5.15% to 8.56% (RSD < 6.90%). NX-5948 was demonstrated to be stable under the present storage conditions. After validation, the method was successfully applied for the quantification of NX-5948 in beagle dog plasma after oral and intravenous administration of the NX-5948.
研究领域:
药物分析
·
生物制药
·
临床药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2024
·Vol. 29
·期 3
·659
·
IF 4.2
Zedian Li,Liangrui Xiang,Shijia Pan,Dahai Zhu,Shen Li,He Guo
This research constructed a novel O3/CaO2/HCO3−system to degrade antibiotic oxytetracycline (OTC) in water. The results indicated that CaO2andHCO3−addition could promote OTC degradation in an O3system. There is an optimal dosage of CaO2(0.05 g/L) andHCO3−(2.25 mmol/L) that promotes OTC degradation. After 30 min of treatment, approximately 91.5% of the OTC molecules were eliminated in the O3/CaO2/HCO3−system. A higher O3concentration, alkaline condition, and lower OTC concentration were conducive to OTC decomposition. Active substances including ·OH,1O2,·O2−, and ·HCO3−play certain roles in OTC degradation. The production of ·OH followed the order: O3/CaO2/HCO3−> O3/CaO2> O3. Compared to the sole O3system, TOC and COD were easier to remove in the O3/CaO2/HCO3−system. Based on DFT and LC-MS, active species dominant in the degradation pathways of OTC were proposed. Then, an evaluation of the toxic changes in intermediates during OTC degradation was carried out. The feasibility of O3/CaO2/HCO3−for the treatment of other substances, such as bisphenol A, tetracycline, and actual wastewater, was investigated. Finally, the energy efficiency of the O3/CaO2/HCO3−system was calculated and compared with other mainstream processes of OTC degradation. The O3/CaO2/HCO3−system may be considered as an efficient and economical approach for antibiotic destruction.
研究领域:
环境科学
·
化学工程
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Foods
·
2022
·Vol. 11
·期 3
·306
·
IF 5.561
Tingting Yang,Yuhang Hu,Yamei Yan,Wangting Zhou,Guijie Chen,Xiaoxiong Zeng,Youlong Cao
The fruits ofLycium barbarumare rich in flavonoids, which may contribute to the health-promoting function ofLycium barbarum. However, the composition of flavonoids in the fruits ofLycium barbarum(LBFs) has received little attention. Thus, the goal of this work was to identify more kinds of flavonoids from fruits ofLycium barbarumby liquid chromatography–mass spectrometry. The potential antioxidant and anti-inflammatory activities of LBFs in vitro were also investigated. Thirteen flavonoid compounds were identified in LBFs, of which daphnetin, 6,7-dihydroxycoumarin, astragalin, taxifolin, eriodictyol, naringenin, and chrysoeriol were identified for the first time in the fruits ofLycium barbarum, which greatly enriched the variety of flavonoids in the fruits ofLycium barbarum. LBFs showed a similar superior antioxidant activity to vitamin C. Furthermore, LBFs exhibited an anti-inflammatory activity by suppressing the production of nitric oxide and pro-inflammatory cytokines, including tumor necrosis factor-alpha, interleukin-1β, and interleukin-6, in lipopolysaccharide-treated RAW264.7 macrophage cells. This study demonstrated the potential development of LBFs as functional foods.Keywords:Lycium barbarum;flavonoids;liquid chromatography mass spectrometer (LC-MS);antioxidant;anti-inflammatory
研究领域:
药理学
·
生物化学
·
营养学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Applied Sciences-Basel
·
2025
·Vol. 15
·期 3
·1486
·
IF 2.5
Mengye Jia,Riya Jin,Yina Qiao,Yu Jiang,Jiaoqin Liu,Zengdi He,Zhaoguo Liu,Jingshuai Gao
2,2′,4,4′,6,6′-hexanitrostilbene (HNS) is a heat-resistant, low-sensitivity energetic material with widespread applications in pliable linear-shaped charges, high-temperature-resistant oil well perforating charges, and rocket propellants. However, the presence of highly toxic and mutagenic nitroaromatic compounds in HNS wastewater necessitates efficient and accurate detection methods. Unfortunately, the existing analytical methods for HNS detection are outdated and incompatible with modern equipment. This limits their application due to issues with detection range, accuracy, and cost. To address this gap, an improved method was developed using an Ultimate 3000 UHPLC system with methanol and water as the mobile phase and UV-Vis detection at 271 and 226 nm wavelengths. The results indicate that the optimal detection conditions are achieved with a methanol-to-water ratio of 70:30 and a flow rate of 0.5 mL/min, providing high accuracy and efficiency. Compared to traditional methods, this approach reduced the detection time by nearly 70%, with the shortest analysis time ranging from 6 to 6.5 min, significantly lowering the cost of HNS detection. The method demonstrated excellent linearity (R2> 0.9999) and high sensitivity within the concentration range of 0.50–150.00 mg/L, with precise and reliable results. This work provides both theoretical insights and experimental validation for the detection and analysis of HNS in wastewater.
研究领域:
分析化学
·
环境科学
·
含能材料
·
色谱学
✓ 已验证
产品关联
DOI
高影响因子
Frontiers in Molecular Biosciences
·
2023
·
IF 5
Xue Zhichao,Zeng Jiaming,Yin Xinchi,Li Yongshu,Meng Bo,Zhao Yang,Fang Xiang,Gong Xiaoyun,Dai Xinhua
Palbociclib is a specific CDK4/6 inhibitor that has been widely applied in multiple types of tumors. Different from cytotoxic drugs, the anticancer mechanism of palbociclib mainly depends on cell cycle inhibition. Therefore, the resistance mechanism is different. For clinical cancer patients, drug resistance is inevitable for almost all cancer therapies including palbociclib. We have trained palbociclib resistant cells in vitro to simulate the clinical situation and applied LC-MS multi-omics analysis methods including proteomic, metabolomic, and glycoproteomic techniques, to deeply understand the underly mechanism behind the resistance. As a result of proteomic analysis, the resistant cells were found to rely on altered metabolic pathways to keep proliferation. Metabolic processes related to carbohydrates, lipids, DNA, cellular proteins, glucose, and amino acids were observed to be upregulated. Most dramatically, the protein expressions of COX-1 and NDUFB8 have been detected to be significantly overexpressed by proteomic analysis. When a COX-1 inhibitor was hired to combine with palbociclib, a synergistic effect could be obtained, suggesting the altered COX-1 involved metabolic pathway is an important reason for the acquired palbociclib resistance. The KEGG pathway of N-glycan biosynthesis was identified through metabolomics analysis. N-glycoproteomic analysis was therefore included and the global glycosylation was found to be elevated in the palbociclib-resistant cells. Moreover, integration analysis of glycoproteomic data allowed us to detect a lot more proteins that have been glycosylated with low abundances, these proteins were considered to be overwhelmed by those highly abundant proteins during regular proteomic LC-MS detection. These low-abundant proteins are mainly involved in the cellular biology processes of cell migration, the regulation of chemotaxis, as well as the glycoprotein metabolic process which offered us great more details on the roles played by N-glycosylation in drug resistance. Our result also verified that N-glycosylation inhibitors could enhance the cell growth inhibition of palbociclib in resistant cells. The high efficiency of the integrated multi-omics analysis workflow in discovering drug resistance mechanisms paves a new way for drug development. With a clear understanding of the resistance mechanism, new drug targets and drug combinations could be designed to resensitize the resistant tumors.
研究领域:
肿瘤学
·
药理学
·
分子生物学
·
系统生物学
·
生物信息学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2023
·
IF 3.1
Xiaomei Wu,Bin Pang,Jianwei You,Leiye Yu,Ruobing Ren,Hongyang Wang,Li Chen
Sphingosine-1-phosphate (S1P) is an active signaling metabolite synthesized by blood cells, exported into blood stream, and can trigger many downstream signaling pathways with disease implications. Understanding how S1P is transported is of great values for dissecting the function of S1P, but most existing methods for measuring S1P transporter activity use radioactive substrates or involve multiple workup steps, hindering their broader uses. In this study, we develop a workflow combining sensitive LC-MS measurement and a cell-based transporter protein system to measure the export activity of S1P transporter proteins. Our workflow demonstrated good applications in studying different S1P transporters SPNS2 and MFSD2B, WT and mutated protein, and different protein substrates. In summary, we provide a simple yet versatile workflow for measuring the export activity of S1P transporters, facilitating future studies of S1P transport mechanism and drug development.
研究领域:
生物化学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）