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Open AccessFeature PaperArticle

Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood

1
Department of Biomedical Sciences, School of Medicine, University of Sassari, Sassari 07100, Italy
2
Department of Clinical Pharmacology, College of Medicine and Public Health, Flinders University and Flinders Medical Centre, Adelaide SA 5042, Australia
3
Department of Chemistry and Pharmacy, University of Sassari, Sassari 07100, Italy
4
Quality Control Unit, University Hospital of Sassari (AOU-SS), Sassari 07100, Italy
*
Author to whom correspondence should be addressed.
Molecules 2018, 23(12), 3326; https://doi.org/10.3390/molecules23123326
Received: 23 November 2018 / Revised: 13 December 2018 / Accepted: 13 December 2018 / Published: 14 December 2018
(This article belongs to the Special Issue Anti-Inflammatory and Anti-Allergy Agents in Medicinal Chemistry-II)
Given that the peculiar redox behavior of ergothioneine involves a rapid regeneration process, the measurement of its precursor and redox metabolite hercynine could be particularly useful in assessing its role in oxidative stress or other biological processes. Thus, a LC-MS/MS method for the determination of hercynine concentrations in whole blood was developed. After lysis of red blood cells by cold water, samples were filtered on micro concentrators at a controlled temperature of 4 °C. The clear filtered fluid was then treated with diethylpyrocarbonate to derivatize hercynine for the analysis by LC-MS/MS. The derivatized analyte was isocratically separated as a carbethoxy derivative on a C18 column with a mobile phase of an aqueous 0.1% v/v formic acid and acetonitrile (95:5). Effluents were monitored by MRM transitions at m/z 270.28→95 and 273.21→95 for hercynine and its deuterated counterpart, respectively. No cross-talk between MRM transitions was observed and a good linearity was found within a range of 35–1120 nmol/L. The LOD and LOQ were, respectively, 10.30 and 31.21 nmol/L with an intraday and intermediate precision below 7%. The average hercynine concentration in whole blood from 30 healthy male volunteers (aged 77 ± 12 years) was 178.5 ± 118.1 nmol/L. Overall, the method is easy to perform, allowing a rapid and accurate assessment of whole blood concentrations of hercynine. View Full-Text
Keywords: aminothione; dismutatiion; LC-tandem mass spectrometry; antioxidant; zwitterion aminothione; dismutatiion; LC-tandem mass spectrometry; antioxidant; zwitterion
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Sotgia, S.; Murphy, R.B.; Zinellu, A.; Elliot, D.; Paliogiannis, P.; Pinna, G.A.; Carru, C.; Mangoni, A.A. Development of an LC–Tandem Mass Spectrometry Method for the Quantitative Analysis of Hercynine in Human Whole Blood. Molecules 2018, 23, 3326.

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