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Headspace solid-phase microextraction – gas chromatography – mass spectrometry for the quantitative determination of the characteristic flavouring agent eugenol in serum samples after enzymatic cleavage to validate post-offence alcohol drinking claims

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CONTRIBUTORS:
  Author Schulz, Katja
  Author Schlenz, Katja
  Author Malt, Steffen
  Author Metasch, Robert
  Author Römhild, Wolfgang
  Author Dreßler, Jan
  Author Lachenmeier, Dirk W (Chemisches und Veterinäruntersuchungsamt Karlsruhe)
JOURNAL:
  Journal of Chromatography A, 1211(1-2), 113 - 119.
YEAR: 2008
PUB TYPE: Journal Article
SUBJECT(S): Alcoholic beverages; Flavouring agents; Congener analysis; Beverage-characteristic aroma compounds; Eugenol; Bitter; HS-SPME–GC–MS; Metabolism
DISCIPLINE: Medicine
HTTP: http://dx.doi.org/10.1016/j.chroma.2008.09.068
LANGUAGE: English
PUB ID: 103-445-512 (Last edited on 2008/11/03 03:28:45 US/Mountain)
SPONSOR(S):
 
ABSTRACT:
A rapid headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME–GC–MS) method has been developed for the determination of eugenol in serum samples after enzymatic cleavage. Eugenol is a characteristic marker for the consumption of certain alcoholic beverages including some digestif bitters and herbal liqueurs as well as wood-cask-aged spirits. This method enables the detection of eugenol with a limit of detection (LOD) of 3.2 ng/ml and a limit of quantification (LOQ) of 4.8 ng/ml in serum samples with excellent precision (5.3% intraday, 6.9% interday) and linearity (correlation coefficient R2 = 0.992). Our findings confirm that eugenol undergoes a rapid phase II metabolism as it occurs completely conjugated as eugenol glucuronide in serum. Free eugenol was not detectable in any of our samples, which necessitated enzymatic cleavage with â-glucuronidase prior to HS-SPME sampling. In vivo experiments were conducted with a volunteer, who consumed a digestif bitter beverage on three different days under controlled conditions. At defined intervals, blood samples were taken from the subject. Using these blood samples, concentration/time profiles for serum eugenol glucuronide were determined. A rapid resorption leads to a peak eugenol glucuronide concentration directly after drinking (up to 1742 ng/ml if 78 mg of eugenol are ingested) followed by a decrease during the next 3 h. Blood samples were also taken from 20 drivers claiming to have consumed drinks containing eugenol. In five of the samples, eugenol glucuronide was detected at serum concentrations ranging from 12.1 to 172.3 ng/ml. These test results, in particular, confirm that the analysis of volatile compounds can be useful in forensic toxicology for the verification of post-offence alcohol consumption claims.
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