Use of modified Doehlert‐type experimental design in optimization of a hybrid electrospray ionization ion trap time‐of‐flight mass spectrometry technique for glutathione determination

Zachariadis, George A.; Rosenberg, Erwin

doi:10.1002/rcm.6475

Cited by 13 publications

(5 citation statements)

References 28 publications

(60 reference statements)

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“…However, adjustments of critical MS parameters, such as voltage and temperature, are needed to ensure stable spray and prevent gas-phase fragmentation of glycans as they travel from the emitter, through the S-lens, and into the quadrupole. Although the magnitude of these effects are instrument specific, previous work on mass spectrometer peptide optimization has indicated that the basic relationships behind these parameters are to some extent universal. ,− Despite evidence that design of experiments can lead to improvements in protein abundance, to date, there has been no parallel investigation of MS parameters for glycans.…”

Section: Resultsmentioning

confidence: 99%

Definitive Screening Design Optimization of Mass Spectrometry Parameters for Sensitive Comparison of Filter and Solid Phase Extraction Purified, INLIGHT PlasmaN-Glycans

2015

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High-throughput, quantitative processing of N-linked glycans would facilitate large-scale studies correlating the glycome with disease and open the field to basic and applied researchers. We sought to meet these goals by coupling Filter-Aided-N-Glycan Separation (FANGS) to the individuality normalization when labeling with glycan hydrazide tags (INLIGHT™) for analysis of plasma. A quantitative comparison of this method was conducted against solid phase extraction (SPE), a ubiquitous and trusted method for glycan purification. We demonstrate that FANGS-INLIGHT purification was not significantly different from SPE in terms of glycan abundances, variability, functional classes, or molecular weight distributions. Furthermore, to increase the depth of glycome coverage, we executed a definitive screening design of experiments (DOE) to optimize the MS parameters for glycan analyses. We optimized MS parameters across five N-glycan responses using a standard glycan mixture, translated these to plasma and achieved up to a three-fold increase in ion abundances.

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Section: Resultsmentioning

confidence: 99%

Definitive Screening Design Optimization of Mass Spectrometry Parameters for Sensitive Comparison of Filter and Solid Phase Extraction Purified, INLIGHT PlasmaN-Glycans

2015

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“…GSH is one of the most abundant intracellular antioxidants in animal cells. Since it is considered to be one of the most abundant tripeptides in human organism, and its action against xenobiotics and oxidative radicals is well known (Zachariadis and Rosenberg, 2013). In mice, exposure to TCDD results in significant depletion of GSH by oxidative stress in liver (Slezak et al, 2002).…”

Section: Discussionmentioning

confidence: 99%

Ameliorative effects of docosahexaenoic acid on the toxicity induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in cultured rat hepatocytes

2014

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The 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is an environmental contaminant toxicant that mediates carcinogenic effects associated with oxidative DNA damage. Docosahexaenoic acid (DHA) with antioxidant functions has many biochemical, cellular, and physiological functions for cells. The present study assessed, for the first time, the ameliorative effect of DHA in alleviating the toxicity of TCDD on primary cultured rat hepatocytes (HEPs). In vitro, isolated HEPs were incubated with TCDD (5 and 10 μM) in the presence and absence of DHA (5, 10, and 20 μM) for 48 h. The cell viability was detected by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) release. DNA damage was analyzed by liver micronucleus assay and 8-oxo-2-deoxyguanosine (8-OH-dG) level. In addition, total antioxidant capacity (TAC) and total oxidative stress (TOS) were assessed to determine the oxidative injury in HEPs. The results of MTT and LDH assays showed that TCDD decreased cell viability but not DHA. On the basis of increasing treatment concentrations, the dioxin caused significant increases of micronucleated HEPs and 8-OH-dG as compared to control culture. TCDD also led to significant increases in TOS content. On the contrary, in cultures treated with DHA, the level of TAC was significantly increased during treatment in a concentration-dependent fashion. DHA showed therapeutic potential against TCDD-mediated cell viability and DNA damages. As conclusion, this study provides the first evidence that DHA has protective effects against TCDD toxicity on primary cultured rat hepatocytes.

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“…The methods of detecting biothiols include high performance liquid chromatography (HPLC) [23, 24], ultraviolet visible spectrometry [25, 26], electrochemical method [27, 28], capillary electrophoresis [29], mass spectrometry [30], and so forth. Among them, fluorescence method has attracted much attention due to its high sensitivity and efficiency [31–38]; in particular it can be used for real-time imaging of living tissues by means of confocal detection [39–41].…”

Section: Introductionmentioning

confidence: 99%

A New Quinone Based Fluorescent Probe for High Sensitive and Selective Detection of Biothiols and Its Application in Living Cell Imaging

Liu

Wong

et al. 2019

International Journal of Analytical Chemistry

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In view of the vital role of biothiols in many physiological processes, the development of simple and efficient probe for the detection of biothiols is of great medical significance. In this work, we demonstrate the use of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), which respond rapidly to biothiols especially to glutathione, as a new fluorescent probe for the selective detection and bioimaging of biothiols. This new fluorescent probe can distinguish glutathione from cysteine and homocysteine easily under physiological concentration and detect glutathione quickly within three minutes. This probe exhibits high selectivity to biothiols and the detection limit was determined to be 3.08 × 10−9 M for glutathione, 8.55 × 10−8 M for cysteine, and 2.17 × 10−9 M for homocysteine, respectively. The sensing mechanism was further explored by density functional theory (DFT) and nuclear magnetic resonance (NMR) experiment; results showed that the interaction forces between the probe and biothiols were electrostatic interaction. In addition, the probe has been successfully applied to the detection of biothiols in Eca9706 cells by fluorescence confocal imaging technology.

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Use of modified Doehlert‐type experimental design in optimization of a hybrid electrospray ionization ion trap time‐of‐flight mass spectrometry technique for glutathione determination

Abstract: The results highlighted the possibilities offered by the employment of multifactorial optimization towards the improvement of performance of tandem mass spectrometry techniques.

Cited by 13 publications

References 28 publications

Definitive Screening Design Optimization of Mass Spectrometry Parameters for Sensitive Comparison of Filter and Solid Phase Extraction Purified, INLIGHT PlasmaN-Glycans

Definitive Screening Design Optimization of Mass Spectrometry Parameters for Sensitive Comparison of Filter and Solid Phase Extraction Purified, INLIGHT PlasmaN-Glycans

Ameliorative effects of docosahexaenoic acid on the toxicity induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin in cultured rat hepatocytes

A New Quinone Based Fluorescent Probe for High Sensitive and Selective Detection of Biothiols and Its Application in Living Cell Imaging

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