查看更多>>摘要：Rationale SCO-267 is a potent full agonist of G-protein-coupled receptor 40. As a promising therapeutic agent for type 2 diabetes mellitus, it is necessary to elucidate its metabolite profiles during the stage of drug development for safety considerations. Methods The in vitro metabolism was investigated by incubating SCO-267 (5 mu M) with liver microsomes and hepatocytes (rat and human). For in vivo metabolism, SCO-267 (10 mg/kg) was orally administered to rats and plasma samples were collected. The metabolites were identified via measurements of accurate mass, elemental composition and product ions using liquid chromatography coupled to hybrid quadrupole Orbitrap high-resolution mass spectrometry (LC-Orbitrap-MS). Results A total of 19 metabolites were structurally identified. M2 (hydroxyl-SCO-267), M15 (SCO-267-acyl-glucuronide), M16 (desmethyl-SCO-267) and M17 (desneopentyl-SCO-267) were verified with reference standards. M2, M11, M16 and M17 were the major metabolites originating from hydroxylation, O-demethylation and N-dealkylation, respectively. Phenotyping study with recombinant human P450 enzymes demonstrated that hydroxylation (M2 and M11) was mainly catalyzed by CYP2C8 and 3A4; demethylation (M16) was mainly catalyzed by CYP2D6, and less catalyzed by CYP2C8 and 3A4; and N-dealkylation (M17) was exclusively triggered by CYP3A4. Conclusions Hydroxylation, O-demethylation, N-dealkylation and acyl glucuronidation were the major metabolic pathways of SCO-267. This study is the first to discover the metabolic fates of SCO-267, which provides a basis for safety assessment of this drug candidate.
查看更多>>摘要：Rationale Fully formulated oils (FFOs) are integral to automotive lubrication; however, detailed compositional analysis is challenging due to high levels of chemical complexity. In particular, existing mass spectrometric approaches often target particular FFO components, leading to poor analytical coverage of the overall formulation, with increased overheads and analytical timescales. Methods Herein we report the application of a commercially available SICRIT SC-20 dielectric barrier discharge ionisation (DBDI) source and Thermo Fisher Scientific LTQ Orbitrap XL to the analysis of an FFO. Nitrogen was used as a discharge gas for the DBDI source, and was modified using a range of commonplace solvents to tailor the experimental conditions for the analysis of various components. Results The reported method allowed analysis of a range of FFO components of interest, encompassing a wide range of chemistries, in under 1 min. By modifying the discharge gas used for ionisation, experiments could be optimised for the analysis of particular FFO components across positive and negative ion modes. In particular, use of water vapour as a discharge gas modifier with positive ion mode mass spectrometry permitted concomitant analysis of antioxidants and base oil hydrocarbons. Furthermore, case studies of selected linear alkanes and alkenes profile the differences in the range of ions formed across these saturated and unsaturated aliphatic compounds, giving insight into the fate of base oil hydrocarbons in FFO analyses. Conclusions A rapid method for analysis of FFO compositions has been developed and provides coverage of a range of components of interest. The results indicate that the method presented may be of utility in analysis of other FFOs or similarly challenging complex mixtures.
查看更多>>摘要：Rationale Polyhalogenated carbazoles (PHCZs) are dioxin-like compounds that are ubiquitous in the environment. However, their unintentional emissions from industrial sources have received little attention and there is no method available for determination of PHCZs in industrial waste. This research develops a method for determination of PHCZs in industrial waste. Methods In this research, a glass column packed with activated silica serves as a rapid and efficient clean-up pretreatment for purification. An isotope dilution gas chromatography/triple quadrupole tandem mass spectrometry method was established for simultaneous determination of eleven PHCZs in industrial samples. Results The regression coefficients of the standard curves for the congeners were all >0.99. The method detection limit ranged from 1.46 to 3.82 ng/mL for liquid samples and from 0.009 to 0.021 ng/g for solid samples. The precision described by the relative standard deviation ranged from 2.4% to 18.4% for liquid samples and from 5.5% to 35.8% for solid samples. The recovery ranges for the liquid and solid samples were 82%-123% and 83%-137%, respectively. 3-Chlorocarbazole (3-CCZ) and 36-dichlorocarbazole (36-CCZ) can be detected in both chemical bottom liquid from vinyl chloride production and fly ash from medical waste incineration by this method. Conclusions An efficient method is established for determination of PHCZs from industrial waste. The discovery of 3-CCZ and 36-CCZ highlights the importance of identification of potential industrial sources of PHCZs and clarification of their contribution to environmental risks. Our method could be applied to investigate industrial emission of PHCZs.
查看更多>>摘要：Rationale: Embedding resins are commonly used to facilitate high-resolution sampling for stable isotope analysis but anomalous delta C-13 values have been observed in some cases. Here we compare the results of microsampling strategies for hand-drilled versus resin-embedded micromilled samples from the same marine shells to assess whether resin contamination is implicated in delta C-13 spikes. The comparison allows assessment of the relative benefits for spatial resolution, seasonal range for both delta O-13 and delta C-13, and sample failure rates. Methods: Hand-drilled samples were obtained from two bivalve shells (Spisula sachalinensis), corresponding to micromilled samples on the same shells where high delta C-13 spikes were observed. All carbonate powders were analysed using a dual-inlet Isoprime mass spectrometer and Multiprep device. Results from both sample sets were compared statistically. Results: No anomalous high delta C-13 values and no failures due to insufficient gas were observed in the hand-drilled samples in contrast to the embedded micromilled sequences. Spatial resolution was reduced (similar to 2.5 x) in the former compared with the latter, resulting in a small reduction in the total range observed in the micromilled delta C-13 and delta O-13 values. Reduced sampling resolution between the two datasets was only significant for delta O-13. Conclusions: For S. sachalinensis (as with other similar bivalves), rapid growth mitigates the reduced sampling resolution of hand drilling and does not significantly impact observed isotopic range and seasonal patterning. Occurrence of anomalous delta C-13 values were eliminated and failure rates due to insufficient sample size greatly reduced in the hand-drilled dataset. We can find no other explanation for the occurrence of delta C-13 spikes than contamination by the embedding agent. We conclude that the logistical and interpretational benefits of careful hand drilling may be preferable to resin embedding for micromilling in marine shells, corals or speleothems where growth rate is rapid and the highest resolution is not required.
查看更多>>摘要：Rationale Poly- and perfluoroalkyl substances (PFASs) and organophosphorus flame retardants (OPFRs) are two types of emerging organic pollutants with potential human health hazards. Here, a rapid and sensitive method was developed for the determination of sixteen PFASs and seven OPFRs in human serum by ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS). Methods After optimizing the chromatographic and mass spectrometric conditions, respectively, 100 mu L of serum sample was liquid-liquid extracted using 8 mL of methyl tert-butyl ether. The 23 targets were quantified within 8 min. All targets were quantified by the isotope-internal standard method in both negative- and positive-ion mode by UPLC/MS/MS. Results The method was validated in terms of sensitivity, linearity, precision, and accuracy. The limit of detection ranged between 0.004 and 0.650 ng/mL. Recoveries ranged from70.0% to 118.9% with a relative standard deviation lower than 20%. The developed method was successfully applied to analyze targeted analytes in human serum samples. A total of 13 of 23 analytes were detected in over 50% of samples. Conclusions A sensitive and rapid method was developed to quantify sixteen PFASs and seven OPFRs in serum. Sensitivity, linearity, recovery, and precision were validated and found to be satisfactory. This method can be a valuable tool for evaluation of exposure to both PFASs and OPFRs with high separation efficiency and sensitivity.
查看更多>>摘要：Rationale Cucumber, as a popular fruit and vegetable, has tremendously contributed to providing a sufficient and high-quality food supply. However, the cucumber plant metabolites, which may possess potential benefits for human health, were rarely reported. In addition, rapid detection of these metabolites from the complex biological matrix of cucumber samples is a tremendous challenge. Methods A rapid detection method was established to systematically screen cucurbitacins and cucurbitacin glycosides in cucumber plants by combining high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC-Q-TOF-MS) with in-source fragmentation (ISF). Moreover, the alkali cations, including acetic acid, 0.1% LiCl, 0.1% NH4Cl, 0.1% NaCl, and 0.1% KCl, were added to the mobile phase for improving the ion response. Results The fragmentation pathways of seven cucurbitacins and cucurbitacin glycosides were primarily investigated. The characteristic ISF ions at m/z 501.3211 and 503.3367 were identified and employed to screen 40 cucurbitacins and cucurbitacin glycosides from the complex biological matrix. Their structures were identified by their tandem mass spectrometry (MS/MS) spectra and fragmentation pathways of references. Finally, the metabolic distribution and network of cucurbitacins and cucurbitacin glycosides in cucumber plants were also proposed. Conclusions This work marks the first systematic and comprehensive study of the metabolites in cucumber plants using HPLC-Q-TOF-MS technology, providing a template for screening and identifying the triterpenoids from other plant-derived medicines or food.
查看更多>>摘要：Rationale We show evidence of cysteinylation on Cys252 of recombinant human p40 subunit of interleukin 12 (IL-12). This was reported in 1996. However, no paper detailing this concept has been published yet. Our paper reports the quantification of Cys252 cysteinylation as well as the full disulfide bridges assignment by nonreducing peptide mapping using mass spectrometry (MS) detection. Methods Nonreducing peptide mapping was applied for disulfide bridges assignment. This study presents an ad hoc method in which applying a neutral pH in the presence of an alkylating agent allowed to mitigate the formation of artifacts such as reshuffled disulfide bridges and permitted the detection of free cysteine. Ultra-high-performance liquid chromatography-MS analysis was performed on a Waters quadrupole time-of-flight Xevo G2-XS mass spectrometer acquiring data in MSE mode. MS data were processed using Expressionist MS Refiner 13.5 (Genedata). Results Scouting experiments were performed using two batches of drug substance. An in-depth study of the LC tandem mass spectrometry profiles revealed the presence of additional species related to "free" Cys252; this cysteine residue was also detected in its S-cysteinylated and S-homocysteinylated forms. This result is consistent with that reported in literature so far. The relative abundance of overall "cysteinylated" species resulted in the range between 46% and 36%, which has also been confirmed using orthogonal techniques such as Ellman's assay. Conclusions Our data clearly demonstrate that the free cysteine (Cys252) on the p40 subunit of recombinant IL-12 is also present in its cysteinylated and homocysteinylated forms at a considerable rate. Our observations, although based on results obtained on an IL-12-derived fusion protein, are consistent with the current literature.
NSTL
Wiley