JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2026
·
IF 3.4
Kun Yang,Jianhua Wu,Chengming Yang,Liang Liu
A Protein G magnetic bead-based LC-MS/MS method was developed for dupilumab quantification in human serum. • The surrogate peptide SGQSPQLLIYLGSNR enabled sensitive and specific targeted detection (LLOQ 1.00 µg/mL). • Excellent linearity, precision and accuracy were achieved across a wide dynamic range (1.00–400 µg/mL). • Considerable interpatient variability was observed in clinical trough samples. Dupilumab, a monoclonal antibody targeting IL-4Rα, is used for the treatment of type 2 inflammatory diseases. Recent studies have revealed a nonlinear relationship between its serum concentration and clinical response, particularly a “biological tipping point” during dose interval prolongation, highlighting the need for individualized therapeutic drug monitoring (TDM). In this study, a robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for dupilumab quantification in human serum was developed and validated. Specifically, dupilumab was enriched from 50 µL serum using Protein G magnetic beads, followed by denaturation, reduction, alkylation, and tryptic digestion. A surrogate peptide SGQSPQLLIYLGSNR was selected for quantification, and its stable isotope-labeled (SIL) analog served as an internal standard. Separation was performed on an Acquity UPLC BEH C 8 column (50 mm × 2.1 mm, 1.7 µm) with detection using positive electrospray ionization in multiple reaction monitoring (MRM) mode. Excellent linearity was achieved from 1.00 to 400 µg/mL (r² > 0.99), with a lower limit of quantification of 1.00 µg/mL. Intra- and inter-run precision (%CV) and accuracy (%Bias) were within 9.27%. The validated method was successfully applied to analyze clinical serum samples. The established LC-MS/MS method provides a reliable alternative to immunoassays, with a wide dynamic range suitable for TDM and personalized dose interval adjustments in patients receiving dupilumab. Download: Download high-res image (93KB) Download: Download full-size image
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DOI
高影响因子
Frontiers in Chemistry
·
2024
·
IF 3.8
Baodong Feng,Linqi Su,Yang Yang,Renyan Liu,Yu Zhang,Lingyi Xin,Li Wang,Zhiming Yang,Xuemei Wei,Qinhua Chen
Background: Ulcerative colitis (UC) is a chronic inflammatory bowel disease (IBD) influenced by multiple factors. Berberine, an isoquinoline alkaloid derived from the root and bark of Coptis chinensis Franch., has shown promise in managing UC, but its underlying mechanisms remain unclear.Methods: To elucidate the relationship between berberine, ulcerative colitis (UC), and the organism’s metabolome, we established a dextran sulfate sodium (DSS)-induced UC model in rats. Colonic tissue was collected for histopathological examination, while plasma samples were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with dynamic Multiple Reaction Monitoring (dMRM). This approach, characterized by its short analysis time of 20 min per sample, excellent reproducibility, and straightforward data processing, allowed for the comprehensive detection of a wide array of metabolites, including amino acids, lipids, and organic acids, many of which are implicated in the pathophysiology of UC.Results: Our results showed that berberine modulated the metabolic disturbances of 33 compounds in the plasma of UC rats, primarily including amino acids, pyrimidines, organic phosphoric acids, fatty acyls, and organonitrogen compounds. These altered metabolites were associated with various pathways, such as amino acid metabolism, glutathione metabolism, nicotinate and nicotinamide metabolism, taurine and hypotaurine metabolism, pyrimidine metabolism, glyoxylate and dicarboxylate metabolism, and the citrate cycle (TCA cycle). Notably, 3-hydroxyproline, homocysteic acid, L-threonine, L-lysine, carbamoyl phosphate, O-phosphoethanolamine, taurine, leucine, and phosphorylcholine exhibited significant differences between the Treatment and Model groups, with levels reverting to those of the Control group (p < 0.001). These findings suggested that these compounds may serve as potential plasma biomarkers for UC.Conclusion: This study provided valuable insights into the mechanism by which berberine exerted its therapeutic effects on UC through metabolomics. Our results highlighted berberine’s potential to modulate key metabolic pathways and restore the levels of several metabolites, suggesting its utility as a therapeutic agent for UC. These findings underscored the importance of metabolomics in understanding the pathophysiology and treatment of UC.
研究领域:
药理学
·
代谢组学
·
炎症性肠病研究
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DOI
PubMed
高影响因子
XENOBIOTICA
·
2025
·
IF 1.2
Shanshan Chen,Donghan Wang,Yuanyuan Zhao,Yaqi Sun,Yueyao Luan,Qixuan Sun,Jiaqi Wang,Yuhang Yan,Jing Yu,Chunhua Zhou
With the increasing prevalence and escalating complexity of mental disorders, precise medication has become critically important. This necessitates an efficient, accurate, and convenient method for drug concentration monitoring to support laboratory personnel and clinicians. In this study, three liquid chromatography-tandem mass spectrometry methods were developed and validated for simultaneously determining and quantifying 27 antipsychotics and related metabolites in human plasma. The plasma samples were subjected to protein precipitation using methanol, with isotope-labelled internal standards (ISs), followed by separation via isocratic elution on a BEH C18 column. Mass spectrometric analysis was performed using electrospray ionisation in positive ionisation mode with multiple reaction monitoring for quantitative detection. The analytes demonstrated high separation efficiency, with a single sample run time of 3.0 min. The method exhibited a wide linear range with excellent linearity across the concentration range. The intra- and inter-batch precision were ≤10.00%, the accuracy was 88.67–113.29%. Accurate quantification of antipsychotics remained unaffected under various storage conditions: 72 h at room temperature, 7 d at 4 °C refrigeration, and 14 d at −80 °C freezing. This validated methodology has been successfully applied to plasma samples from patients with psychiatric disorders, demonstrating its practical utility for accurate quantification of antipsychotics in large-scale and complex matrices containing multiple analytes.
研究领域:
临床医学
·
分析化学
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DOI
PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
·
2024
·
IF 4
Can Cui,Xiaojun Jiao,Xianbin Xu,Jiongzhou Gong,Jiaxin Zhou,Ge Tian,Liwei Lu,Li Li,Xianhui Huang
The presence of residues of veterinary drugs in eggs is an ongoing and significant issue of concern. Fluralaner is widely used in the prevention of ectoparasites, including the poultry red mite, but no analytical method has been reported for detection of its residues in eggs. An HPLC-MS/MS method was established for the determination of fluralaner residues in whole eggs and validated for LOD, LOQ, specificity, linearity, accuracy, precision, and matrix effects. LOD was 5 μg•kg −1 and LOQ was 65 μg•kg −1 . Recovery ranged from 97.7% to 103.7% (spiked at 65, 650, 1300, and 2600 μg•kg −1 ), while intra-assay and inter-assay coefficients of variation were consistently below 1.5%. A drug withdrawal trial was conducted with 16 laying hens administered a therapeutic dose of fluralaner in drinking water on days 0 and 7, with eggs being analyzed for 23 days. The highest concentration, 1235 μg•kg −1 , was detected on day 13. This was below the European Union maximum residue limit for fluralaner in edible chicken tissues and eggs (1300 μg•kg −1 ). Residues were detectable up to two weeks after cessation of treatment. This method is suitable for routine compliance monitoring of fluralaner residues in egg matrices and may be adapted for analysis of other edible tissues.
研究领域:
兽医学
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食品科学
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DOI
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2021
·Vol. 44
·期 13
·2510-2523
·
IF 3.645
Zili Guo,Peixi Zhu,Xiaoai He,Tianhe Yan,Xianrui Liang
A valid and reliable method based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry using electrospray ionization was established to identify chemical constituents in the ethanol extract of pigeon pea leaves. A total of 58 compounds were detected both in positive and negative modes. Among them, 42 compounds, including 16 flavones, 1 flavonol, 5 flavanones, 9 isoflavones, 1 coumarin, 1 lactone, 6 stilbenes, 2 chalcones, and 1 other compound, were unambiguously identified or tentatively assigned in view of the retention time, the molecular formula, as well as the fragmentation patterns. Moreover, eight sets of isomers were differentiated by the ion trap mass spectrometry based on the fragment ion differences or the abundance differences of the same fragment ions. The energy-resolved mass spectrometry in light of the relative abundance of characteristic fragment ions was adopted in the study.
研究领域:
植物化学
·
分析化学
·
药用植物研究
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DOI
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2024
·
IF 3.1
Mengru Bai,Qian Shen,Yong Wu,Zhiyuan Ma,Yuqing Wang,Mingyang Chen,Dan Liu,Lin Zhou
Methotrexate (MTX) is commonly prescribed as the initial treatment for gestational trophoblastic neoplasia (GTN), but MTX monotherapy may not be effective for high-risk GTN and choriocarcinoma. The cellular uptake of MTX is essential for its pharmacological activity. Thus, our study aimed to investigate the cellular pharmacokinetics and transport mechanisms of MTX in choriocarcinoma cells. For the quantification of MTX concentrations in cellular matrix, a liquid chromatography-tandem mass spectrometry method was created and confirmed initially. MTX accumulation in BeWo, JEG-3, and JAR cells was minimal. Additionally, the mRNA levels of folate receptor α (FRα) and breast cancer resistance protein (BCRP) were relatively high in the three choriocarcinoma cell lines, whereas proton-coupled folate transporter (PCFT), reduced folate carrier (RFC), and organic anion transporter (OAT) 4 were low. Furthermore, the expression of other transporters was either very low or undetectable. Notably, the application of inhibitors and small interfering RNAs (siRNAs) targeting FRα, RFC, and PCFT led to a notable decrease in the accumulation of MTX in BeWo cells. Conversely, the co-administration of multidrug resistance protein 1 (MDR1) and BCRP inhibitors increased MTX accumulation. In addition, inhibitors of OATs and organic-anion transporting polypeptides (OATPs) reduced MTX accumulation, while peptide transporter inhibitors had no effect. Results from siRNA knockdown experiments and transporter overexpression cell models indicated that MTX was not a substrate of nucleoside transporters. In conclusion, the results indicate that FRα and multiple transporters such as PCFT, RFC, OAT4, and OATPs are likely involved in the uptake of MTX, whereas MDR1 and BCRP are implicated in the efflux of MTX from choriocarcinoma cells. These results have implications for predicting transporter-mediated drug interactions and offer potential directions for further research on enhancing MTX sensitivity.
研究领域:
药理学
·
肿瘤学
·
细胞生物学
·
分析化学
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DOI
PubMed
高影响因子
Frontiers in Pharmacology
·
2022
·
IF 5.988
Hu Yan,Wang Zhe,Liu Jiayue,Yang Wen,Yang Qiang,Liu Yuan-Cai,You Qiu-Yun,Chen Xiao-Jia,Wan Jian-Bo
As a prevalent medicinal liquor among Chinese people, a type of Chinese herbal spirit from Jing Brand Co., Ltd (CHS-J) is a newly developed health beverage with the health functions of anti-fatigue and immune enhancement. The researchers from the enterprise found that the contents of several components in CHS-J samples have been significantly decreasing during the stated storage period, as detected by the HPLC-UV method, which would make a great challenge for quality control of CHS-J. Furthermore, the chemical stability of CHS-J during the storage period is greatly challenged affected, especially in the environment of high temperature and light exposure. To systematically reveal the unstable components and promote the quality control of CHS-J, the chemical stability of CHS-J during the shelving storage period was characterized by the UPLC/Q-TOFMS-based metabolomics approach. First, the targeted and untargeted metabolomics approaches discovered the significantly changed components in CHS-J samples produced in different years. Furthermore, the accelerated tests of newly produced CHS samples and several authorized standards were conducted to validate the above results and elucidate the possible mechanisms underlying these chemical changes. Moreover, these chemical changes during the storage period had little influence on the anti-fatigue effect of CHS-J samples. These findings will offer new insight into the understanding of the chemical stability of CHS-J and will facilitate the quality control of CHS-J.
研究领域:
食品科学
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中药学
·
分析化学
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DOI
PubMed
高影响因子
CHEMICAL RESEARCH IN TOXICOLOGY
·
2022
·
IF 3.973
Liming Wang,Shuo Huang,Tongtong Zhu,Xiaoyan Ge,Chenxi Pei,Ge Hong,Lifeng Han
Iohexol, the raw material of nonionic X-ray computed tomography (X-CT) contrast medium, is usually injected into the vein before CT angiography diagnosis. It is used for angiography, urography, and lymphography. With the advantages of low contrast density and good tolerance, it is currently one of the most popular contrast media. However, the renal toxicity of iohexol seriously affects its safety use. Therefore, it is of great importance to identify new potential diagnostic biomarkers and therapeutic targets in the process of contrast medium-induced acute kidney injury (CI-AKI) in order to safely use iohexol in clinical practice. In this study, in order to understand the metabolic mechanism of CI-AKI, ultra-high-performance liquid chromatography/quadrupole-Orbitrap-mass spectrometry and 1H NMR-based metabolomic techniques were utilized to study the metabolic spectra of kidney, plasma, and urine from CI-AKI rats, and a total of 30 metabolites that were closely related to kidney injury were screened out, which were mainly related to 9 metabolic pathways. The results further indicated that iohexol might intensify kidney dysfunction in vivo by disrupting the metabolic pathways in the body, especially through blocking energy metabolism, amino acid metabolism, and promoting inflammatory reactions.
研究领域:
药理学
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毒理学
·
代谢组学
·
临床医学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Chemistry
·
2023
·
IF 5.5
Ma Yufeng,Zhang Fengsen,Zhong Yijing,Huang Yongchun,Yixizhuoma,Jia Qiangqiang,Zhang Shoude
Phosphodiesterase type 5 (PDE5), a cyclic nucleotide phosphodiesterase, controls the duration of the cyclic guanosine monophosphate (cGMP) signal by hydrolyzing cGMP to GMP. Inhibiting the activity of PDE5A has proven to be an effective strategy for treating pulmonary arterial hypertension and erectile dysfunction. Current enzymatic activity assay methods for PDE5A mainly use fluorescent or isotope-labeled substrates, which are expensive and inconvenient. Here, we developed an LC/MS-based enzymatic activity assay for PDE5A without labeling, which detects the enzymatic activity of PDE5A by quantifying the substrate cGMP and product GMP at a concentration of 100 nM. The accuracy of this method was verified by a fluorescently labeled substrate. Moreover, a new inhibitor of PDE5A was identified by this method and virtual screening. It inhibited PDE5A with an IC50 value of 870 nM. Overall, the proposed strategy provides a new method for screening PDE5A inhibitors.
研究领域:
药理学
·
生物化学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2023
·Vol. 107
·期 10
·3217-3227
·
IF 5
Wang Yang,Chen Huanhuan,Zhao Meiqi,Feng Lei,Liu Zexin,Zeng Qiongya,Shi Wenqiang,Zhu Wen,Song Luyao,Zhu Jianwei,Lu Huili
Abstract Being an important immune stimulant of T lymphocytes and NK cells, the recombinant human interleukin-15 (rhIL-15) has been extensively researched in tumor immunotherapy or as a vaccine adjuvant. However, the rhIL-15 manufacturing level lags far behind its growing clinical demand due to the lack of efficient and exact analysis methodologies to characterize the trace by-products, typically redox and deamidation. In order to improve the production and quality control of rhIL-15, here we developed an expanded resolution reverse-phase high-performance liquid chromatography (ExRP-HPLC) approach to quickly and accurately analyze the oxidation and reduction by-products of rhIL-15, which may appear during the purification processes. Firstly, we developed RP-HPLC methods which can separate rhIL-15 fractions with different levels of oxidization or reduction, respectively, and the redox status of each peak was then determined by measuring the intact mass with a high-resolution mass spectrometer (UPLC-MS). To further clarify the complex pattern of oxidization of specific residues, the peaks with various oxidation levels were digested into pieces for peptide mapping to pinpoint the exact changes of oxygen and hydrogen atoms in the rhIL-15 by-products. In addition, we performed the ExRP-HPLC and UPLC-MS analysis of partially deamidated rhIL-15 to characterize their oxidation and reduction. Our work is the first in-depth characterization of the redox by-products of rhIL-15, even for deamidated impurities. The ExRP-HPLC method we reported can facilitate the rapid and accurate quality analysis of rhIL-15, which is substantially helpful for streamlining the industrial manufacturing of rhIL-15 to better meet the demands of clinical applications. Keypoints • The oxidization and reduction rhIL-15 by-products were characterized for the first time.
• The changes of oxygen and hydrogen atoms in rhIL-15 redox by-products were accurately determined by UPLC-MS.
• Oxidation and reduction by-products of deamidated rhIL-15 were further analyzed.
研究领域:
生物化学
·
药学
·
免疫学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2023
·Vol. 148
·期 12
·2782-2792
·
IF 4.2
Zhifang Yang,Chenyu Wang,Lan Xiao,Chuang Wang,Li Tang,Lei Guo,Jianwei Xie
The accurate and sensitive detection of active biotoxin proteins and the determination of their kinetics are vital for the upsurge of chemical attacks but still limited. Herein, we report a liquid chromatography-tunable ultraviolet spectroscopic-quadrupole mass spectrometric detection (LC-TUV-QDa) method of active ricin. This method has the advantage of the accurate quantification of active ricin in decreased oligonucleotide (oligo) substrates as well as the produced adenine, in which the QDa detection offers the confirmative evidence of oligo and adenine products. We invented a strong cation exchange (SCX)-tip sample pretreatment way to facilitate the requirement of clean product injection without any fouling proteins. After full-method validation, a wide linear range of 1–5000 ng mL−1 was obtained with a high sensitivity of 1 ng mL−1 active ricin based on the most preferable deoxynucleobase-hybrid RNA (Rd) substrate, Rd12, and without any enrichment. We also fully depicted the kinetic parameters of ricin and its six Rd or RNA substrates and evaluated 11 nucleobase-modified oligos as substrates based on Rd12. Further, we fulfilled an improved molecular docking analysis and revealed that the binding of Rd12 to ricin was more likely to occur at pH 7.4 (typical in vitro and in vivo conditions) than at pH 4.0 (typical ex vitro conditions). With the aid of SCX-tip as a microenzymatic reactor, we can exert the catalytic activity of ricin as N-glycosidase in pH 7.4 toward its Rd12 substrate, with a comparable catalytic efficiency at pH 4.0. This is the first successful implementation of an ex vitro experiment toward oligo substrates at neutral pH, standing on the shoulder of plenty of previously reported efforts all performed under acidic conditions. This method will provide a new and powerful way to detect active ricin when tackling relevant problems in public safety and security.
研究领域:
分析化学
·
毒理学
·
生物技术
·
生物化学
·
公共安全与安保
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产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2015
·Vol. 5
·期 51
·40764-40771
·
IF 3.84
Rong Li,Yuefa Jia,Jun Wu,Qiang Zhen
The degradation behavior of oxytetracycline (OTC) in aqueous solution using Fe2O3–TiO2 nanopowders (Fe2O3–TiO2 NPs) as photocatalysts under UV/visible light was investigated in this paper. Fe2O3–TiO2 NPs photocatalysts synthesized using co-precipitation were characterized by XRD, TEM, FT-IR and UV-vis DRS. Fe2O3–TiO2 NPs with 45 wt% Fe2O3 exhibited higher photocatalytic activity than others. The photocatalytic degradation of OTC shows a maximum efficiency at pH = 5.5 under both UV/visible light due to the surface adsorption. The intermediate products during the photocatalytic degradation of OTC were detected by LC-MS/TOF. Seven main intermediates were formed, and their evolution was discussed. On the basis of the evidence oxidative intermediate formation and a detailed degradation path way of OTC by 45 Fe2O3–TiO2 NPs are proposed.
研究领域:
环境科学
·
化学
·
材料科学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）