scholarly journals Cell-Wide Survey of Amide-Bonded Lysine Modifications by Using Deacetylase CobB

2019 ◽  
Vol 21 (1) ◽  
Author(s):  
Yun Wei ◽  
Wan-Jie Yang ◽  
Qi-Jun Wang ◽  
Peng-Cheng Lin ◽  
Jian-Yuan Zhao ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Protein Function ◽  
Gas Chromatography Mass Spectrometry ◽  
Human Tissues ◽  
Metabolomic Analysis ◽  
Dynamic Changes ◽  
Post Translational Modifications ◽  
Covalently Bonded ◽  
Different Origins

Abstract Background Lysine post-translational modifications are important regulators of protein function. Proteomic and biochemical approaches have resulted in identification of several lysine modifications, including acetylation, crotonylation, and succinylation. Here, we developed an approach for surveying amide-bonded lysine modifications in the proteome of human tissues/cells based on the observation that many lysine modifications are amide-bonded and that the Salmonella enterica deacetylase, CobB, is an amidase. Results After the proteome of human tissues/cells was denatured and the non-covalently bonded metabolites were removed by acetone washes, and the amide-bonded modifiers were released by CobB and analyzed using liquid- and/or gas chromatography/mass spectrometry metabolomic analysis. This protocol, which required 3–4 days for completion, was used to qualitatively identify more than 40 documented and unreported lysine modifications from the human proteome and to quantitatively analyze dynamic changes in targeted amide-bonded lysine modifications. Conclusions We developed a method that was capable of monitoring and quantifying amide-bonded lysine modifications in cells of different origins.

scholarly journals Metabolomic Analysis of Gingival Crevicular Fluid Using Gas Chromatography/Mass Spectrometry

2016 ◽  
Vol 5 (1) ◽  
pp. A0047-A0047 ◽  
Author(s):  
Miho Ozeki ◽  
Takenori Nozaki ◽  
Jun Aoki ◽  
Takeshi Bamba ◽  
Kirk R. Jensen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Gingival Crevicular Fluid ◽  
Gas Chromatography Mass Spectrometry ◽  
Metabolomic Analysis ◽  
Chromatography Mass Spectrometry ◽  
Crevicular Fluid

Occurrence of Organochlorine Pesticides in Human Tissues Assessed Using a Microextraction Procedure and Gas Chromatography–Mass Spectrometry

2020 ◽  
Author(s):  
Marta Pastor Belda ◽  
José Antonio González-Franco ◽  
Ricardo Rubio ◽  
Natalia Campillo ◽  
Manuel Hernández-Córdoba ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Organochlorine Pesticides ◽  
Abdominal Fat ◽  
Gas Chromatography Mass Spectrometry ◽  
Human Tissues ◽  
Salting Out ◽  
Chromatography Mass Spectrometry ◽  
Relative Standard ◽  
Dispersive Liquid Liquid Microextraction

Abstract This work focuses on the development, validation and application of an analytical method for the determination of twenty organochlorine pesticides (OCPs) in human tissues using salting-out liquid–liquid extraction and dispersive liquid–liquid microextraction for sample preparation and gas chromatography–mass spectrometry to analyze the obtained extracts. Measurement of the concentration levels of these toxics in tissues can be used to assess the risk of the population to exposure. The linearity of the proposed method was verified in the 10–1,000 ng/g range. The sensitivity was evaluated calculating the limits of detection (LODs) for 20 OCPs (α-, β-, γ- and δ-hexachlorocyclohexane (HCH), α- and β-endosulfan, endosulfan sulfate, aldrin, dieldrin, endrin, endrin ketone, endrin aldehyde, α- and γ-chlordane, 4,4′-dichlorodiphenyltrichloroethane, 4,4′-dichlorodiphenyldichloroethylene (DDE), 4,4′-dichlorodiphenyldichloroethane, heptachlor, heptachlor epoxide and methoxychlor), most of them being found between 1.0 and 16 ng/g. The intra- and interday precisions were <12% for relative standard deviation values. The accuracy of the method was evaluated by recovery studies, which gave recovery percentages in the 85–109% range. Seven different tissues (liver, kidney, heart, spleen, lung, brain and abdominal fat) from eight autopsies were analyzed, and only three cases were seen to have β-HCH and 4,4′-DDE in abdominal fat, while 4,4′-DDE was also detected in the heart of one case. The rest of the samples were free of the studied OCPs at least above the corresponding LODs.

scholarly journals Drug Analysis in Necrophagous Flies and Human Tissues

2007 ◽  
Vol 58 (3) ◽  
pp. 313-316 ◽  
Author(s):  
Marija Definis-Gojanović ◽  
Davorka Sutlović ◽  
Dolores Britvić ◽  
Bože Kokan
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Quantitative Analysis ◽  
Cause Of Death ◽  
Human Tissue ◽  
Drug Analysis ◽  
The Body ◽  
Gas Chromatography Mass Spectrometry ◽  
Human Tissues ◽  
Multiple Samples

Drug Analysis in Necrophagous Flies and Human TissuesNecrophagous insects may provide useful information about the time, place and cause of death. In addition, they can serve as reliable alternative specimens for toxicological analysis in cases where human tissue and fluids, normally taken during autopsies, are not available, due to decomposition of the corpse. This paper reports the results of drug analysis of the larvae of two fly families, Calliphoridae and Sarcophagidae, collected from the body of a middle-aged man who had committed suicide approximately three weeks before his corpse was found. Multiple samples of decomposed human tissue, of the blowfly, and of the larval flesh were analysed using gas chromatography/mass spectrometry (GC/MS), and amphetamine was detected in all samples. While the screening results were beyond doubt, the quantitative analysis was less clear, and further research is needed in this area.

scholarly journals Identification and Quality Evaluation of Raw and Processed Asarum Species Using Microscopy, DNA Barcoding, and Gas Chromatography-Mass Spectrometry

2020 ◽  
Vol 2020 ◽  
pp. 1-12
Author(s):  
Guangzhe Yao ◽  
Wenjuan Ma ◽  
Xuhua Huang ◽  
Qi Jia ◽  
Jiayuan Shen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Dna Barcoding ◽  
Gas Chromatography Mass Spectrometry ◽  
Volatile Components ◽  
Processing Methods ◽  
Chromatography Mass Spectrometry ◽  
Microscopic Features ◽  
Bioactive Ingredients ◽  
Different Origins

Asarum (Aristolochiaceae) is one of the common herbs used to relieve exterior syndromes. Some volatile components of Asarum which have toxic effect may cause adverse reactions such as headache, general tension, unconsciousness, and respiratory paralysis. Therefore, Asarum is normally processed to reduce such toxicity and adverse effects. The bioactive ingredients contained in different Asarum herbs vary significantly; this variation may be attributed to their differences in species, origins, or processing methods. In this study, 16 batches of Asarum herbs were collected, and their species were identified using DNA barcoding, which is a method for distinguishing plant species, coupled with microscopy. A gas chromatography-mass spectrometry (GC-MS) method for simultaneous determination of 10 compounds was established to evaluate the contents of raw and processed Asarum herbs. Multivariate analysis was then applied to compare different batches of herbs based on the GC-MS data. DNA barcoding identified the herbs as being derived from four sources, and herbs from different origins showed different microscopic features. The results demonstrated that most of the samples were clearly clustered into distinct groups that corresponded to species types. All raw and processed samples were classified by partial least squares discriminant analysis (PLS-DA) based on the 10 analyzed compounds. The findings suggested that safrole and methyleugenol with a variable importance in the project (VIP) > 1 are unique compounds that can be used to differentiate between Asarum species. Safrole, methyleugenol, and 2,6,6-trimethylcyclohepta-2,4-dien-1-one were identified as significant constituents, the presence of which can be used to differentiate between raw and processed Asarum samples. These results indicate that species and processing methods show important effects on the composition of Asarum herbs.

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