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Journal of chromatography
Elsevier
Journal of chromatography

Elsevier

1570-0232

Journal of chromatography/Journal Journal of chromatography
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    UPLC-MS/MS method for the quantification of MCI-77, a novel sigma-1 receptor ligand, and its application to pharmacokinetic studies

    Popa, RalucaKamble, Shyam H.Kanumuri, Raju S.King, Tamara I....
    7页
    查看更多>>摘要：Sigma-1 receptors are involved in pain modulation, particularly in cases of nerve injury and neuropathic pain. High-affinity ligands with improved pharmacokinetic profiles are needed to further investigate the properties of these receptors and their potential as a therapeutic target. The novel compound MCI-77 is one such selective sigma-1 receptor ligand, and the purpose of this study was to characterize its preclinical pharmacokinetic parameters. An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated to quantify MCI-77 in mouse plasma and brain homogenate. The method was validated for sensitivity, selectivity, linearity, accuracy, precision, stability, and dilution integrity. The method has a linearity range of 2-200 ng/mL, a short run-time of 3.2 min, and requires a low sample volume of 25 mu L. A simple protein precipitation procedure was used for compound extraction. Samples were run on an Acquity UPLC BEH C-18 column (1.7 mu m, 2.1 x 50 mm) following a gradient elution method using a mobile phase consisting of water containing 0.1% (v/v) formic acid and acetonitrile. The method was applied to the analysis of plasma and brain homogenate samples from preclinical pharmacokinetic studies in CD-1 mice. MCI-77 exhibited high systemic clearance (8.5 +/- 0.3 L/h/kg) and extensive tissue distribution indicated by a high volume of distribution (20.1 +/- 0.3 L/kg). The concentration levels were consistently higher in brain samples than in plasma (brain/plasma AUC ratio 2.9), indicating its ability to cross the blood-brain barrier.
      原文链接:
    • NSTL
    • Elsevier

    Revealing therapeutic targets and mechanism of baicalin for anti-chronic gastritis using proteomic analysis of the gastric tissue

    Huo, YanZhang, YuWang, XinhongJi, Wanli...
    9页
    查看更多>>摘要：Background: Baicalin (BCL) is a natural compound associated with antioxidant, anti-inflammatory and immunomodulatory activities, among many others. To investigate the therapeutic effect of BCL treatment in ethanolinduced chronic gastritis, we investigated proteomic changes in the gastric tissue to elucidate the therapeutic targets of BCL in chronic gastritis using tandem mass tag (TMT)-based quantitative proteomics. Methods: Sprague-Dawley (SD) rats received 8.7 ml/kg body weight of 56% ethanol intragastrically, which induced chronic gastritis model. Then, BCL (50 mg/kg) or omeprazole (20 mg/kg) was orally administered for 7 days to treat the induced chronic gastritis. After sacrifice, the total protein of the gastric antrum was determined. In addition, a tandem tag labelling for relative and absolute quantification (TMT)-based liquid chromatography with tandem mass spectrometry analysis was performed on the sample to identify differentially expressed proteins (DEPs) between therapy and model groups. Furthermore, the potential protein targets, signalling pathways and protein-protein interaction (PPI) network were analysed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG). Finally, some potential targets were validated using Western blotting. Results: A total of 4,452 proteins were identified and quantified in the gastric antrum tissue of SD rats using TMTbased quantitative proteomics. Of these, 107 DEPs, including 44 upregulated and 63 downregulated proteins, were discovered in the BCL-treated group compared with the untreated group with ethanol-induced gastritis, of these were 33 callback proteins. Biological information analysis demonstrated that the selected differential proteins were involved in the enriched pathways, including MAPK, PI3K-Akt and NF-kappa B. Furthermore, the expression of TPM2, GIMAP4, ICAM-1 and Mpc1 was validated using Western blotting, which was consistent with the proteomics results. The study results confirmed the reliability of the proteomic analysis. Additionally, BCL could decrease the production of interleukin (IL)-2, IL-8 and tumour necrosis factor-alpha while increasing the expression of epidermal growth factor and B-cell lymphoma-2. Notably, PPI network analysis revealed widespread interactions mediated by BCL. Conclusions: We investigated the effects and potential mechanism of BCL in chronic gastritis. Proteomic technology was used to explore BCL-affected proteins and some signalling pathways. The results may provide important insights into discovering potential target proteins for treating chronic gastritis.
      原文链接:
    • NSTL
    • Elsevier

    Rapid determination of hydrogen sulfide-related metabolites in human urine by UHPLC-ESI-MS/MS with fluorinated ion-pairing reagents

    Lajin, BassamObermayer-Pietsch, BarbaraGoessler, Walter
    5页
    查看更多>>摘要：Although primarily considered as a toxic gas, hydrogen sulfide is naturally produced in humans from the amino acid cysteine in low quantities and is commonly referred to as the "third gaseous signaling molecule ". There is a need to establish reliable biomarkers for exposure to hydrogen sulfide in human matrices not only at toxic levels but also at trace levels. Herein, we report optimized analytical methods for the direct determination of three compounds related to hydrogen sulfide metabolism in human urine without sample preparation; namely, trimethylsulfonium, thiosulfate, and cystine. The methods were based on ion-pair ultra-high performance liquid chromatography (UHPLC) with an anionic fluorinated acid or a cationic fluoroalkylamine, a member of a new generation of ion-pairing reagents, and detection by the electrospray ionization tandem mass spectrometry (ESIMS/MS). Rapid chromatographic separation was achieved in 2-3 min with limits of detection of 0.1, 2.5, and 50 nmol L-1 (nM) for trimethylsulfonium, cystine, and thiosulfate, respectively. Isotopically labeled internal standards were used for each analyte to account for matrix effects and qualifier ions were employed to ensure selectivity. The method was validated for recovery (generally within 80-120%) and repeatability (RSD% 1.0-10%), and applied to investigate the normal concentrations of the three analytes in forty morning first-pass urine samples of healthy volunteers.
      原文链接:
    • NSTL
    • Elsevier

    Simultaneous LC-MS/MS quantification of glucocorticoids, melatonin and its metabolites in hair

    Zhu, MinhuiYuan, LinWu, YanChu, Liuxi...
    15页
    查看更多>>摘要：Background: The long-term sleep state has an important influence on one's physical and mental health. Melatonin (MEL) and cortisol with circadian rhythm are deemed to be potential sleep biomarkers. Considering the rapid metabolism of MEL and cortisol, their main metabolites could be alternative indicators showing higher stability and reliability. However, there is short of research developing the method for simultaneous quantification of MEL, cortisol and their metabolites in hair.Objectives: This study aimed to develop a method for the simultaneous quantification of F, MEL and their main metabolites (cortisone; N-acetyl-serotonin, NAS; 6-hydroxymelatonin, 6-O-MEL and 6-sulfatoxymelatonin, S-OMEL) in human hair based on high-performance liquid chromatography tandem mass spectrometry method, and then explore the relationship between the biomarkers' contents and sleep state.Methods: Analytes were extracted from 20-mg hair in 1 mL methanol at about 27 degrees C, and then analyzed in a mobile phase of 95% methanol and 5% 5 mM ammonium acetate, and identified with an electrospray ionization source in positive ion mode. Hair samples closest to the scalp were collected from 65 undergraduates. Sleep state was measured based on participants' scores of the Pittsburgh Sleep Quality Index, the Epworth Sleepiness Scale and the Morningness/Eveningness Questionnaire.Results: The method showed good linearity with the square of correlation coefficient > 0.99 at the ranges of 0.1-1000 pg/mg for MEL, 0.4-1000 pg/mg for NAS, 1.0-1000 pg/mg for 6-O-MEL, 1.0-1000 pg/mg for S-OMEL, 0.5-1000 pg/mg for cortisol and 1.0-1000 pg/mg for cortisone. It showed the limit of detection ranged from 0.05 to 0.3 pg/mg and the limit of quantification ranged between 0.1 and 1.0 pg/mg for the six analytes. The inter- and intra-day coefficients of variation were < 20%. The compounds could be detected in natural hair samples except for S-O-MEL. The average concentration was 0.18 pg/mg for MEL, 3.5 pg/mg for NAS, 3.8 pg/mg for 6-O-MEL, 20.0 pg/mg for cortisone and 2.8 pg/mg for cortisol. The population analysis revealed that there was positive association between hair cortisone and sleep quality.Conclusions: This study had developed an LC-MS/MS method for simultaneous quantification of MEL, NAS, 6-OMEL, cortisone and cortisol in human hair. Hair cortisone might be a promising biomarker of long-term sleep state.
      原文链接:
    • NSTL
    • Elsevier

    Analysis of plasma free amino acids in diabetic rat and the intervention of Ginkgo biloba leaves extract using hydrophilic interaction liquid chromatography coupled with tandem mass-spectrometry

    Zhang, FanLi, Ding-xiangLu, Dong-yuLu, Yi-fan...
    10页
    查看更多>>摘要：Amino acids (AAs) are important metabolites that are related with diabetes. However, their roles in the initiation and development of diabetes mellitus (DM), especially in the treatment of Ginkgo biloba leaves extract (GBE) have not been fully explored. Thus, we investigated the roles that AAs played in the progression and GBE supplementation of DM rat induced by streptozotocin. The rats were randomly divided into a normal control group treated with drug-free solution, a normal control group treated with GBE, a DM group treated with drug-free solution, and DM group treated with GBE; and maintained on this protocol for 9 weeks. Rat plasma was collected from the sixth week to the ninth week and then analyzed with the optimized hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry method. A total of 17 AAs with differential levels were monitored to indicate dysfunction of AAs metabolism to confirm the occurrence and development of DM. Treatment with GBE partially reversed the changes seen in seven AAs including leucine, isoleucine, tyrosine, glutamic acid, asparagines, lysine and alanine in DM rats, indicating that GBE could prevent the occurrence and development of DM by acting on AAs metabolism. The improvement of those AAs metabolism disorders may play a considerable role in the treatment of GBE on the occurrence and development of DM. Those findings potentially promote the understanding of the pathogenic progression of DM and reveal the therapeutic mechanism of GBE against DM.
      原文链接:
    • NSTL
    • Elsevier

    Optimized sample pre-treatment procedure for the simultaneous UPLC-MS/ MS quantification of ipilimumab, nivolumab, and pembrolizumab in human serum

    Jong, Karen A. M. deRosing, HildeHuitema, Alwin D. R.Beijnen, Jos H....
    9页
    查看更多>>摘要：Ipilimumab, nivolumab, and pembrolizumab are immune checkpoint inhibiting monoclonal antibodies. Their efficacy has been proven to be correlated with clearance, and hence, bioanalytical assays to study their pharmacokinetics are of pivotal importance. We present the first kit-free sample pre-treatment procedure of only three hours for the Ultra-Performance Liquid Chromatography - tandem Mass Spectrometry (UPLC-MS/MS) simultaneous quantification of ipilimumab, nivolumab, and pembrolizumab in human serum. The conventional bottom-up sample pre-treatment steps for protein MS bioanalysis including pre-digestion purification, denaturation, reduction, alkylation, and digestion were optimized in terms of sensitivity and reproducibility. In the final, optimal sample pre-treatment procedure, samples were purified by protein precipitation with saturated ammonium sulfate solution, reduced with dithiothreitol, denatured with methanol, and digested with trypsin. The method was then validated according to European Medicines Agency (EMA) and the United States Food and Drug Administration (FDA) guidelines for bioanalytical method validation, and 4-6-20 acceptance criteria were applied. This method was selective, accurate, and precise within the range of 3-200 mu g/mL for all analytes. The validated developed assay was applied to determine ipilimumab, nivolumab, and pembrolizumab concentrations in patient serum, and the results were compared to enzyme-linked immunosorbent assay (ELISA) results.
      原文链接:
    • NSTL
    • Elsevier

    Magnetic solid phase extraction followed by in-situ derivatization with core-shell structured magnetic graphene oxide nanocomposite for the accurate quantification of free testosterone and free androstenedione in human serum

    Zhang, XianhuaXu, HuiyuZhou, CongyaYang, Li...
    10页
    查看更多>>摘要：Circulating free androgens are important indicators for a variety of diseases, so accurate determination of these hormones in serum is of great clinical significance. However, there are still many challenges for the accurate quantification of free androgens using mass methods because of their very low levels and complex interferences in serum, as well as the high serum protein binding rate and high nonspecific binding (NSB) rate. Here, an HPLC-MS/MS method coupled with magnetic solid phase extraction (MSPE) and in-situ derivatization was developed to quantify two free androgens-free testosterone (FT) and free androstenedione (FA4)-in human serum simultaneously and accurately. Ultrafiltration was used to obtain free androgens in serum. To minimize the NSB rate and obtain accurate results, the ultrafiltration membrane was doubly modified with surfactant followed by a silane-coupling agent. Multiple pre-saturation with the tested samples was also used. With these strategies, the ultrafiltration recoveries were up to 95.4% for FT and 94.0% for FA4, so the NSB was negligible. After that, the extremely low levels of free androgens in ultrafiltrates were extracted and enriched using MSPE with core-shell structured ferroferric oxide coated with graphene oxide. Hydroxylamine hydrochloride was used to derivatize the analytes and the reaction took place on the surface of the adsorbent. All the extraction and derivatization conditions were optimized. Under such conditions, the assays were linear for FT within the range of 2-100 pg mL-1 and for FA4 within the range of 5-500 pg mL-1. The intra-and inter-run CV was less than 12.3% and 10.9% for FT and less than 7.2% and 8.3% for FA4, respectively. For the intra-and inter-run accuracies, the relative error of the mean was 9.9% and 8.4% for FT, and 11.5% and 7.3% for FA4, respectively. The total extraction recoveries with MSPE in-situ derivatization were 93.2% for FT, 93.8% for FA4 and 95.7% for the internal standard. The method was validated and was used to quantify the trace level of these two free androgens in serum samples from female patients suspected of having polycystic ovary syndrome (PCOS) accurately. It is expected to improve the diagnosis accuracy of PCOS when combined with other clinical indicators.
      原文链接:
    • NSTL
    • Elsevier