ANALYTICAL BIOCHEMISTRY
·
2021
·
IF 3.365
Yan-Ying Xu,Fei Qin,Yi-Xin Dai,Xue-Feng Zhong,Juan Yan,Su-Mei Xu,Ping-Sheng Xu
This study aimed to develop an LC-MS/MS method for determining sildenafil and its metabolites N -desmethylsildenafil and N1,N4 -desmethylsildenafil in human plasma and applying it to a pharmacokinetic study of sildenafil in healthy volunteers. Sildenafil-d8 was used as the internal standard. Plasma samples were pretreated via protein precipitation with acetonitrile . The extractives were then separated on an ACQUITY UPLC BEH C18 (50-mm × 2.1-mm, 1.7-μm) column using gradient elution . The aqueous and organic mobile phases were ammonium formate 2 mM supplemented with 0.1% formic acid in water and acetonitrile, respectively, and the flow rate was 0.3 mL/min. An electrospray ionization source was applied, and multiple reaction monitoring was operated in the positive mode with selective channels at m / z 475.30 → 100.10, 461.20 → 283.30, 483.30 → 108.10, and 449.00 → 283.00 for sildenafil, sildenafil-d8, N - desmethylsildenafil, and N1,N4 -desmethylsildenafil, respectively. The linear calibration curves of sildenafil and its metabolites spanned 1.0–1000 ng/mL. The lower limit of quantification was 1.0 ng/mL. The extractive recovery of analytes from the biological matrix was more than 90% and the matrix effect complied with relevant provisions. The intra- and inter-day precisions of sildenafil and its metabolite were <10%. The intra- and inter-day accuracy of sildenafil, N - desmethylsildenafil, and N1,N4 -desmethylsildenafil was more than 99%. The method is highly sensitive and selective, and it was successfully applied to the bioequivalence studies of 100-mg sildenafil citrate tablets in 40 healthy Chinese volunteers.
研究领域:
药物分析
·
临床药理学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2023
·
IF 3.4
Si-Tao Xu,Hao-Wen Jin,Xin Jin,Bi-Xin Xu,Yu Zhang,Tao Xie,Guangji Wang,Jiankun Wang,Le Zhen
VK2809 is a promising drug candidate in Phase II clinical trials for the treatment of non-alcoholic steatohepatitis (NASH). It is a prodrug with a HepDirect strategy, which can achieve selective hepatic metabolic activation , generating an active metabolite VK2809A as a potent and selective agonist for thyroid hormone receptor beta (TRβ), a concomitant reactive metabolite VK2809B, and a glutathione (GSH) conjugate MB06588. Currently, there is no convenient and sensitive bioanalytical method for the simultaneous determination of the above three metabolites. Herein, we established an LC-MS/MS method to separate VK2809 and its metabolites on the XSelect HSS T3 column and quantified them in negative electrospray ionization mode. Subsequently, several factors were investigated such as the use of 60% acetonitrile for homogenization to stabilize the analytes, the addition of 20 mM glutathione for the derivation of VK2809B, and the protein precipitation with methanol containing Sobetirome as the internal standard (IS). The method exhibited good linearity for all compounds (19.4–388.4 nM for VK2809; 27.4–2744.4 nM for VK2809A and 10.6–211.0 nM for MB06588) with great correlation coefficients (r > 0.996). The method validation also demonstrated acceptable precision (RSD < 13.0% for VK2809, RSD < 7.9% for VK2809A, RSD < 14.4% for MB06588) and accuracy (92.7%−103% for VK2809, 91.2%−107.3% for VK2809A, 96%−106.7% for MB06588). The matrix effect, recovery, and stability were also suitable to determine all the analytes. This method is suitable for the bioanalysis of VK2809 and its metabolites and has been successfully applied to the study of intrahepatic exposure in rats. It is expected to be further practiced in drug design , optimization, and metabolism study in the following research.
研究领域:
药理学
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分析化学
·
生物化学
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
·
2025
·Vol. 15
·期 1
·1-8
·
IF 3.9
Wang Yueyuan,Diao Zhipeng,Qu Yuchen,Fan Kai,Feng Chen,Lv Bo,Yu Yunli,Geng Zhou
Omadacycline, a third-generation tetracycline antibiotic, exhibits time-dependent pharmacokinetics. The ratio of the 24-hour area under the concentration-time curve to minimum inhibitory concentration (AUC 0−24 /MIC) serves as the primary pharmacokinetic/pharmacodynamic (PK/PD) index for omadacycline, demonstrating strong correlation with therapeutic efficacy, particularly in critically ill patient populations. To facilitate routine therapeutic drug monitoring (TDM) in clinical practice, a high-performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS) method was developed for quantification of omadacycline in human plasma. An Agilent 1260 series liquid chromatograph and an API 4000 triple tandem quadrupole mass spectrometer were used for the determination of omadacycline and endocannabinoids. The separation was performed on a Phenomenex KINETEX XB-C18 column (2.6 μm, 3 × 50 mm) with 0.1% formic acid-water and pure acetonitrile as the mobile phases. The LC-MS/MS separation was carried out using a gradient elution procedure at a flow rate of 0.4 mL/ min, and the total run time was 5 min. An electrospray ionization (ESI) was selected, and the detection ions of omadacycline and the internal standard (fexofenadine-d6) were determined by mass spectrometry scanning with multiple reaction monitoring (MRM) in positive ion mode as follows: m/z 557.4→453.4 and m/z 508.4→472.8, respectively. The established linear range (20-2000 ng/mL) effectively covers the plasma concentration range encountered in > 98% of clinical samples, making it suitable for routine therapeutic drug monitoring. The method demonstrated acceptable selectivity, recovery, and matrix effect. The results of intra-day precision and inter-day precision show that the relative standard deviation (RSD) is less than 10%, while the relative error (RE) is within ± 10.00%. In this study, we developed and validated a simple, sensitive and accurate method to quantify the concentration of omadacycline in human plasma using LC-MS/MS. This offers essential technical support for the rational clinical application of omadacycline.
研究领域:
药理学
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分析化学
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临床药理学
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质谱分析
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治疗药物监测
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2021
·
IF 3.935
Mei Qu,Peng Xue,Qi Zhang,Tiantian Lu,Kun Liu,Bingying Hu,Jingjing Pang,Qianqian Xiao,Tongxin Xu,Quande Wang,Zhongzhe Cheng
Solasodine , a major ingredient in Solanaceae family, has various biological functions such as inducing neurogenesis, anticonvulsant and anti-tumor. Its risk assessment has also drawn public attention. However, little is known about its oral bioavailability and metabolic process. In this study, an liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the quantification of solasodine in mice dried blood spot (DBS) samples. To block nonspecific adsorption, DBS samples were pretreated with bovine serum albumin (BSA) and then extracted with ethyl acetate . This method was applied to a pharmacokinetic and bioavailability study of solasodine. The absolute bioavailability was only 1.28%. Thereafter, its metabolites in mice were characterized using an ultra-performance liquid chromatography Q-Exactive high-resolution mass spectrometer (UHPLC-QE-HRMS). Several isomeric metabolites were well separated and differentiated using their retention time, fragmentation pathways and correspondingly fragmentation rules of solasodine. As a result, 21 metabolites were characterized including 16 phase I and 5 phase II metabolites. The proposed metabolic pathways showed that solasodine mainly experienced oxidation, dehydration, dehydrogenation and sulfation. These results could help us to better understand the efficacy and safety of solasodine.
研究领域:
药理学
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代谢组学
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Plant Science
·
2024
·
IF 4.1
Yang Wenjuan,Tang Sidi,Xu Rubing,Zhang Lu,Zhou Zihao,Yang Yong,Li Yanyan,Xiang Haibo
Fusarium is a soil-borne pathogen that poses a serious threat to the quality and yield of hundreds of crops worldwide, particularly tobacco production. Using metabolomics technology, we investigated natural metabolites from disease-conducting soil (DCS) and disease-suppressing soil (DSS) of tobacco rhizosphere as fungicides to control tobacco Fusarium wilt (TFW), which is mainly caused by Fusarium oxysporum. Furthermore, the antifungal mechanisms of these natural metabolites were preliminarily elucidated through various assessments, including antifungal activity determination, chemotaxis effect tests, PI staining experiments, and measurements of extracellular conductivity and protein content. Metabolomics results showed that the DCS with three different disease grades (G1, G5 and G9 groups) had significantly higher levels of 15, 14 and 233 differential rhizosphere metabolites (DRMs) and significantly lower levels of 72, 152 and 170 DRMs compared to the DSS (G0 group). According to KEGG pathway analysis, these DRMs were found to be enriched in the caffeine metabolism, biosynthesis of phenylpropanoids, galactose metabolism and tyrosine metabolism, etc. Linustatin, scopoletin and phenylpropiolic acid were picked out from these DRMs and found to have suppressive activity against F. oxysporum through correlation analysis and antifungal experiments. The three DRMs showed strong inhibitory effects on the growth and spore germination of F. oxysporum at concentrations of 0.5 mM or higher in each test period. Furthermore, F. oxysporum showed a phobotaxis effect against these three DRMs at concentrations as low as 0.25 mM. Finally, we found that the three DRMs had an inhibitory effect on F. oxysporum by destroying the integrity of the cell membrane and increasing the membrane permeability of F. oxysporum. This study firstly reports the inhibition activity of phenylpropiolic acid and linustatin on F. oxysporum, providing a practical and environmentally friendly method for biocontrol of TFW by using natural fungicides.
研究领域:
植物病理学
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代谢组学
·
农业科学
·
真菌学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food Analytical Methods
·
2025
·
IF 3
Lin Congcong,Zhu Xuena,Liu Rui,Zhang Li,Yu Tingting,Lu Qun
The consumption of honey contaminated with phytotoxins, particularly from Ranunculaceae plants, is a global public health concern due to their widespread distribution and diverse toxic compounds. This study developed a UPLC-QQQ-MS method to simultaneously quantify 11 phytotoxins of Ranunculaceae family in honey. Analytical (mobile phase) and pre-treatment (extraction solvent and cleanup column) procedures were optimized, identifying Methanol–Water (added 0.1% formic acid and 5 mM ammonium formate), acetonitrile, and SCX column as optimal. Method validation demonstrated negligible matrix effects, good linearity (R 2 > 0.995), good recoveries (81.38–116.85%), and high precision (RSD ≤ 8.63%) for 25–500 ng/mL ranges. LODs and LOQs were 0.02–0.15 µg/kg and 0.07–0.50 µg/kg, respectively (excluding anemonin). Retrospective analysis of 50 honey samples detected magnoflorine, talatisamine, hypaconitine, senecionine, and protopine at positive rates of 2%, 2%, 2%, 8%, and 10%, respectively. Thus, this work provides important technical supports for the risk assessment of phytotoxins in honey.
研究领域:
分析化学
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食品安全
·
毒理学
·
植物化学
✓ 已验证
产品关联
DOI
高影响因子
CHROMATOGRAPHIA
·
2022
·Vol. 85
·期 3
·221-231
·
IF 2.213
Sun Zhiping,He Lingli,Yang Qingqing,Zhang Haizhi,Xu Weiren,Qin Xinguang,Liu Gang,Hu Zhongze,Zhang Luyong,Liu Changxiao
The influence of genetic polymorphism of metabolic enzymes on drug–drug interactions (DDI) should be thoroughly investigated owing to its remarkable effect on therapeutic treatments that are futile. Here, different from earlier articles focused on various CYP subfamilies and their isoforms, the effect of genetic polymorphs of a particular isoform, CYP2C9, was comprehensively studied. Using diclofenac as probe substrate, the influence of three CYP2C9 variants (CYP2C9*1, *2 and *3) on DDI was conducted from the measurement of inhibitory ability of six drugs towards three variants. A modified LC–MS/MS method according to former report was constructed for the determination of 4ʹ-hydroxydiclofenac in CYP2C9 enzyme incubation system and validated for accuracy, precision, linearity range within the acceptance criteria for regulatory guidelines. After selecting appropriate incubation time and enzyme concentration, the kinetic parameters of three CYP2C9 variants towards diclofenac were examined and half inhibitory concentrations (IC 50 ) of six drugs towards three CYP2C9 variants were determined. IC 50 of paroxetine and losartan towards CYP2C9*2 were almost threefold higher and 2.5-fold lower than that towards CYP2C9*1, respectively. IC 50 of glibenclamide towards CYP2C9*3 was twofold more than that towards CYP2C9*1. Our results could offer effective guidelines for co-administration of these six drugs with various CYP2C9 substrates in individuals carrying variant CYP2C9 alleles, also broaden the application area of LC–MS/MS.
研究领域:
药理学
·
临床医学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
Applied Food Research
·
2025
·
IF 6.2
Guodong Shan,Xiuqin Fan,Qingcheng Wang,Li Zhang,Lijun Ge,Weibo Lu,Jing Xue,Kang Chen,Zejun Wang,Qing Shen
The inappropriate use of H2 receptor antagonists (H2RAs) in functional foods (FFs) and food-medicine homology materials (FMHs) designed to enhance gastrointestinal function (IGF) presents health risks due to potential adverse effects. In this study, an analytical method that integrates Captiva EMR-Lipid (CEL) solid-phase extraction (SPE) with liquid chromatography tandem mass spectrometry (LC-MS/MS) has been developed for the detection of H2RAs (nizatidine, famotidine, cimetidine, ranitidine, and lafutidine) in these products. Through the optimization of CEL-SPE parameters, such as using 200 mg of sorbent and eluting with ACN/water (80/20, v/v), the recovery rates of H2RAs in IGF-FF and IGF-FMH ranged from 87.4 % to 94.6 % and 90.2 % to 97.1 %, respectively. The assessment of matrix effects revealed that the five H2RAs exhibit matrix suppression, necessitating the construction of matrix-matched standard curves with correlation coefficients (r 2 ) exceeding 0.9991. The detection limits ranged from 0.4 to 0.8 μg·kg −1 and the quantification limits ranged from 1.3 to 2.7 μg·kg −1 , with recovery rates spanning from 85.4 % to 97.1 % and relative standard deviations ≤ 15 %. The successful development of this technique not only helps safeguard consumer health but also provides scientific basis and technical support for market supervision, thereby preventing and addressing food safety risks.
研究领域:
食品安全
·
分析化学
·
药物检测
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2021
·
IF 3.935
Xinying He,Han Cao,Xia Li,Yan Li,Yunqiu Yu
Lysophosphatidylcholine (LPC) was commonly known as a class of significant differential metabolites of high relevance with many diseases including psoriasis, of which the accurate determination is of great importance to diagnosis or prediction to many diseases. However, it is challenging and complicated because of the enormous biological sample complexity and impurities interference. In this study, we synthesized a magnetic nanocomposite MG@PD@TiO 2 and took advantage of the interactions of Lewis acid−base between the phosphate groups in LPCs and Ti ions on MG@PD@TiO 2 nanomaterials for selective separation and enrichment of LPCs from complex biological matrix. The solid-phase extraction sample pretreatment process by means of MG@PD@TiO 2 nanomaterials coupled with LC–MS/MS method was then applied to actual determination of six typical LPCs (LPC 10:0, 14:0, 16:0, 18:0, 18:1, 22:0) in human plasma. The extraction conditions were scientifically optimized by single-factor test (adsorbent amount, adsorption and desorption time, elution solvent type, eluant volume). Under the optimal conditions, the detection limits (LOD, S/N = 3) and quantification limits (LOQ, S/N = 10) were 1 and 5 ng/mL for LPC 10:0 and LPC 14:0, 0.02 and 0.1 ng/mL for LPC 16:0 and LPC 18:1, 0.05 and 0.2 ng/mL LPC 18:0 and LPC 22:0, respectively. The intra- and inter-day precisions were 3.82–12.60 % (n = 6) and 3.29–13.50 % (n = 6) respectively, the recoveries were in the range of 91.92–113.69 % and the stability of the analytes in the matrix performed well with RSDs≤15.51 %. Finally, the developed method was successfully applied to the accurate determination of six LPCs biomarkers of plasma in patients with psoriasis (n = 10) and control groups (n = 10).
研究领域:
分析化学
·
医学
·
生物标志物研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2020
·
IF 3.209
Xiangling Feng,Yan Cao,Yufeng Ding,Heng Zheng
Topiroxostat is a recently identified safe and selective xanthine oxidoreductase inhibitor and has been used as a urate-lowing agent for the treatment of hyperuricemia and gout in Japan. In this study, we developed a sensitive, selective and high-throughput method that determines topiroxostat in human plasma using high-performance liquid chromatography-tandem mass spectrometry (HPLC–MS/MS) and it was firstly validated. The samples were prepared by protein precipitation with acetonitrile and dilution of the supernatant in water. The chromatographic separation was conducted on an ACE Excel 5 C18-PFP column (2.1 × 50.0 mm) with gradient elution using a mixture of buffer (2 mM ammonium acetate in 0.1 % formic acid) and acetonitrile as the mobile phase at a flow rate of 0.45 mL/min. The total run time lasted 4.0 min. Tandem mass spectrometry, was performed in positive ionization and multiple reaction monitoring (MRM) modes, with the transitions of m/z 249.2→221.1 for topiroxostat and m/z 253.2→225.1 for topiroxostat-d 4 , respectively. The method was validated and the selectivity, carry-over effect, linearity, accuracy, precision, extraction recovery, matrix effect, and stability were acceptable according to the US Food and Drug Administration and the European Medicines Agency regulations. The method was successfully applied to a clinico-pharmacokinetic study involving twelve healthy Chinese subjects under fasting condition.
研究领域:
药理学
·
临床医学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2024
·
IF 3.8
Jiachun Lin,Xue Yang,Anhui Wang,Jinni Yang,Yuan Zheng,Haijuan Dong,Yuan Tian,Zunjian Zhang,Min Wang,Rui Song
Oxysterols and cholesterol precursors are being increasingly investigated in humans and laboratory animals as markers for various diseases in addition to their important functions. However, the quantitative analysis of these bioactive molecules is obstructed by high structural similarity, poor ionization efficiency and low abundance. The current assay methods are still cumbersome to be of practical use, and their applicability in different bio-samples needs to be evaluated and optimized as necessary. In the present work, chromatographic separation conditions were carefully studied to achieve baseline separation of difficult-to-isolate compound pairs. On the other hand, an efficient sample purification method was established for colon tissue samples with good recoveries of sterols, demonstrating negligible autoxidation of cholesterol into oxysterols. The developed UPLC-APCI-MS/MS method was thoroughly validated and applied to measure oxysterols and cholesterol precursors in colon tissue of dextran sulfate sodium (DSS)-induced mouse colitis models, and it is expected to be successfully applied to the quantitative determination of such components in other tissue samples.
研究领域:
分析化学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Proteomics
·
2016
·
IF 3.867
Wen Zhang,Ting Zhong,Yun Chen
In breast cancer, p53 could be functionally compromised by interaction with several proteins. Among those proteins, MDM2 serves as a pivotal negative regulator and counteracts p53 activation. Thus, the ability to quantitatively and accurately monitor the changes in level of p53-MDM2 interaction with disease state can enable an improved understanding of this protein-protein interaction (PPI), provide a better insight into cancer development and allow the emergence of advanced treatments. However, rare studies have evaluated the quantitative extent of PPI including p53-MDM2 interaction so far. In this study, a LC-MS/MS-based targeted proteomics assay was developed and coupled with co-immunoprecipitation (Co-IP) for the quantification of p53-MDM2 complex. A p53 antibody with the epitope residing at 156–214 residues achieved the greatest IP efficiency. 321KPLDGEYFTLQIR333 (p53) and 327ENWLPEDK334 (MDM2) were selected as surrogate peptides in the targeted analysis. Stable isotope-labeled synthetic peptides were used as internal standards. An LOQ (limit of quantification) of 2 ng/mL was obtained. Then, the assay was applied to quantitatively detect total p53, total MDM2 and p53-MDM2 in breast cells and tissue samples. Western blotting was performed for a comparison. Finally, a quantitative time-course analysis in MCF-7 cells with the treatment of nutlin-3 as a PPI inhibitor was also monitored. Biological significance Proteins do not function as single entities but rather as a team player that has to communicate. Protein-protein interaction (PPI), normally by means of non-covalent contact among binary or large protein complex , is essential for many cellular processes including cancer progression. Thus, the ability to quantitatively and accurately monitor the changes in level of PPI with disease state can enable an improved understanding of PPI, provide a better insight into cancer development and allow the emergence of advanced treatments. However, rare studies have evaluated the quantitative extent of PPI so far. The major issue of current available approaches is the trade-off between sensitivity and specificity. Thus, techniques with the ability to quantify PPIs with both high sensitivity (low false-negative rate) and high specificity (low false-positive rate) are eagerly desired. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based targeted proteomics has shown its potential to study biomolecules because of its high sensitivity, high selectivity and wide dynamic range. In this study, we made an effort to develop a LC-MS/MS-based targeted proteomics assay for the quantitative detection of p53-MDM2 interaction in breast cells and tissue samples.
研究领域:
癌症生物学
·
蛋白质组学
·
质谱分析
·
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）