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Characterization of a Hemoglobin Adduct from Ethyl Vinyl Ketone Detected in Human Blood Samples
Version 2 2015-12-17, 10:39
Version 1 2015-11-25, 06:16
journal contribution
posted on 2015-12-17, 10:39 authored by Henrik Carlsson, Hitesh
V. Motwani, Siv Osterman Golkar, Margareta TörnqvistElectrophiles
have the ability to form adducts to nucleophilic
sites in proteins and DNA. Internal exposure to such compounds thus
constitutes a risk for toxic effects. Screening of adducts using mass
spectrometric methods by adductomic approaches offers possibilities
to detect unknown electrophiles present in tissues. Previously, we
employed untargeted adductomics to detect 19 unknown adducts to N-terminal
valine in hemoglobin (Hb) in human blood. This article describes the
characterization of one of these adducts, which was identified as
the adduct from ethyl vinyl ketone (EVK). The mean adduct level was
40 ± 12 pmol/g Hb in 12 human blood samples; adduct levels from
acrylamide (AA) and methyl vinyl ketone (MVK) were quantified for
comparison. Using l-valine p-nitroanilide
(Val-pNA), introduced as a model of the N-terminal
valine, the rate of formation of the EVK adduct was studied, and the
rate constant determined to 200 M–1h–1 at 37 °C. In blood, the reaction rate was too fast to be feasibly
measured, EVK showing a half-life <1 min. Parallel experiments
with AA and MVK showed that the two vinyl ketones react approximately
2 × 103 times faster than AA. The EVK-Hb adduct was
found to be unstable, with a half-life of 7.6 h. From the mean adduct
level measured in human blood, a daily dose (area under the concentration–time-curve, AUC) of 7 nMh EVK was estimated. The AUC of AA from intake via food is about 20 times higher. EVK is naturally
present in a wide range of foods and is also used as a food additive.
Most probably, naturally formed EVK is a major source to observed
adducts. Evaluation of available toxicological data and information
on occurrence of EVK indicate that further studies of EVK are motivated.
This study illustrates a quantitative strategy in the initial evaluation
of the significance of an adduct detected through adduct screening.