Kinetics in serum and urinary excretion of ethyl sulfate and ethyl glucuronide after medium dose ethanol intake

Article (PDF Available)inInternational Journal of Legal Medicine 122(2):123-8 · March 2008with415 Reads
DOI: 10.1007/s00414-007-0180-8 · Source: PubMed
Abstract
The direct ethanol metabolites, ethyl glucuronide (EtG) and ethyl sulfate (EtS), are of increasing importance for clinical and forensic applications, but there are only few studies on the kinetics of EtG in serum and none on EtS. In this study, 13 volunteers (social drinkers) drank ethanol in the form of white wine to reach a blood alcohol concentration of 0.51 +/- 0.17 g/kg, and blood and urine samples were analyzed for EtG and EtS simultaneously by chromatography-tandem mass spectrometry (LC-MS/MS). Mean peak serum EtG and EtS concentrations were 2.9 +/- 1.3 and 2.8 +/- 1.6 micromol/l, respectively, and were reached between 4.0 +/- 0.9 h after the start of drinking (3.0 +/- 0.5 h for EtS). The mean time differences between reaching maximum blood ethanol levels and serum metabolite levels were 2.3 +/- 0.9 h for EtG and 1.2 +/- 0.5 h for EtS. In the last blood samples collected (10-11 h after the start of drinking), 11 (of 13) volunteers were still positive for EtG in serum, whereas only 2 were positive for EtS. In the serum of one female person, no EtS was detectable at any time; however, it was excreted in the urine in (low) concentrations. Ethanol was detectable in the serum for up to 8.6 h after the start of drinking, whereas EtG and EtS were detectable up to more than 5.8 h (EtG) and 4.0 h (EtS), respectively. Mean peak urinary concentrations were 401 +/- 232 micromol/l for EtG and 266 +/- 153 micromol/l for EtS, and mean peak levels were reached 6.2 +/- 0.9 h (EtG) and 5.3 +/- 1.2 h (EtS) after the start of drinking. Maximum concentrations of EtG and EtS in serum showed a wide interindividual variation and could not be correlated to the maximum blood ethanol concentrations. Correlations (p < 0.001, Kendall's Tau b) were found when comparing pairs of parameters, but mostly involved areas under the curve (AUC) of metabolites or of ethanol; one correlation linked the peak concentrations of EtG and EtS in urine.
ORIGINAL ARTICLE
Kinetics in serum and urinary excretion of ethyl sulfate
and ethyl glucuronide after medium dose ethanol intake
Claudia C. Halter & Sebastian Dresen &
Volker Auwaerter & Friedrich M. Wurst &
Wolfgang Weinmann
Received: 7 December 2006 / Accepted: 10 May 2007 /Published online: 9 June 2007
#
Springer-Verlag 2007
Abstract The direct ethanol metaboli tes, ethyl glucuronide
(EtG) and ethyl sulfate (EtS), are of increasing importance
for clinical and forensic applications, but there are only few
studies on the kinetics of EtG in serum and none on EtS. In
this study, 13 volunteers (social drinkers) drank ethanol in
the form of white wine to reach a blood alcohol concentration
of 0.51±0.17 g/kg, and blood and urine samples were an-
alyzed for EtG and EtS simultaneously by chromatography-
tandem mass spectrometry (LC-MS/MS). Mean peak serum
EtG and EtS concentrations were 2.9±1.3 and 2.8±1.6 μmol/l,
respectively, and were reached between 4.0±0.9 h after the
star t of drinking (3.0±0.5 h fo r EtS). The mean time
differences between reaching maximum blood ethanol levels
and serum metabolite levels were 2.3±0.9 h for EtG and 1.2±
0.5 h for EtS. In the last blood samples collected (1011 h
after the start of drinking), 11 (of 13) volunteers were still
positive for EtG in serum, whereas only 2 were positive for
EtS. In the serum of one female person, no EtS was detectable
at any time; however, it was excreted in the urine in (low)
concentrations. Ethanol was detectable in the serum for up to
8.6 h after the start of drinking, whereas EtG and EtS were
detectable up to more than 5.8 h (EtG) and 4.0 h (EtS),
respectively. Mean peak urinary concentrations were 401±
232 μmol/l for EtG and 266±153 μmol/l for EtS, and mean
peak levels were reached 6.2±0.9 h (EtG) and 5.3±1.2 h (EtS)
after the start of drinking. Maximum concentrations of EtG
and EtS in serum showed a wide interindividual variation and
could not be correlated to the maximum blood ethanol
concentrations. Correlations (p<0.001, KendallsTaub)
were found when comparing pairs of parameters, but mostly
involved areas under the curve (AUC) of metabolites or of
ethanol; one correlation linked the peak concentrations of
EtG and EtS in urine.
Keywords Ethyl glucuronide
.
Ethyl sulfate
.
Ethanol markers
.
Kinetics
.
LC-MS/MS
Introduction
Biological markers are becoming more and more important
to prove ethanol consumption due to the growing impact of
alcoholism on modern societies. Most of the ethanol con-
sumed is oxidized to acetaldehyde and acetic acid, but a
small part undergoe s nonoxidative transformation. The
nonoxidative, direct metabolites, ethyl glucuronide (EtG),
ethyl sulfate (EtS), phosphatidyl ethanol (PEth), and fatty
acid ethyl esters (FAEE) can be used as markers for alcohol
consumption besides classical state markers such as
carbohydrate deficient transferrin (CDT), gamma glutamyl
transferase (GGT), or mean corpuscular volume (MCV) [18].
Furthermore, EtG and EtS close the gap in the detection
window between short-term markers (e.g., ethanol) and
long-term markers like CDT, GGT, and MCV. EtG has a far
longer detectability time than ethanol itself [1517], of up
to 80 h in urine. In literature, there has been a discu ssion
Int J Legal Med (2008) 122:123128
DOI 10.1007/s00414-007-0180-8
Electronic supplementary material The online version of this article
(doi:10.1007/s00414-007-0180-8) contains supplementary material,
which is available to authorized users.
C. C. Halter
:
S. Dresen
:
V. Auwaerter
:
W. Weinmann (*)
Institute of Forensic Medicine, University Hospital Freiburg,
Albertstrasse 9,
79104 Freiburg, Germany
e-mail: wolfgang.weinmann@uniklinik-freiburg.de
F. M. Wurst
Psychiatric University Clinic,
Wilhelm-Klein-Strasse 27,
4025 Basel, Switzerland
about stability of ethanol and EtG postmortem and about
generation of ethanol due to putrefaction without generation
of EtG, which can be used for interpretation of postmortem
ethanol concentrations [11]. EtS seems to have a similar
urinary excretion profile as EtG, but further studies with
larger numbers of volunteers are needed to find valid
correlations [3, 6, 10]. Interindividual variations of the
formation of EtG and EtS can be explained by considerable
polymorphisms in the genes coding for enzymes responsi-
ble for EtG and EtS synth esis, namely, uridine diphosphate
(UDP)-glucuronosyltransferases, mainly UGT1A1 and
UGT2B7, [2 , 5] and sulfotransferases [ 12 , 13]. This study
was performed to obtain more knowledge about the
formation and excretion kinetics of both EtG and EtS in
blood and urine.
Materials and methods
Participants and specimen collection
The study was approved by the Ethics Commission of the
University of Freiburg (201/02-05). Informed consent was
obtained before the beginning of the experiments.
For this study, 13 healthy volunteer social drinkers (age
1942 years; six females, seven males; BMI 1829.5 kg/m
2
;
see Table 1) who abstained from alcohol for at least 1 week
gave blood samples 2 days before the experi ment to obtain
normal hemograms [i.e., mean corpuscular volume MCV,
glutamate oxaloacetate transaminase (GOT), glutamate
pyruvate transaminase (GPT), gamma-glutamyl transferase
(GGT), which wer e all within normal physiological limits].
On the first day of the experiment, blank urine and blood
samples were taken before the volunteers were asked to
consume an amount of alcohol, which would lead to a
blood ethanol concentration of 0.50.8 g/kg, calculated by
the Widmark equation corresponding to sex, weight, and
height [1]. The start of drinking was directly after a
standardized breakfast (roll with jam or honey, caffeine-
free tea and coffee) and drinking took about 30 min.
From 15 h after the start of drinking, venous blood
samples were taken at intervals of 30 min, from 510 h at
intervals of 60 min. Urine samples were obtained approx-
imately every 2 h until 10 h after the start of drinking,
followed by three to four samples evenly distributed over
the next day (day 2) and the morning urine of day 3.
Approximately 3.5 and 9.5 h after the start of drinking, the
volunteers were given standardized meals (after 3.5 h pizza
and mineral water, after 9.5 h bread with sausage and
cheese and mineral water). After the last blood sample, the
volunteers were sent home with instructions to abstain from
ethanol contained in drinks, food, or medications until the
end of urine sampling.
Reagents and instrumentation
High performance liquid chromatography (HPLC)-grade
acetonitrile and formic acid (analytical grade) were obtained
from Merck (Darmstadt, Germany). EtG and pentadeuterated
EtG (D
5
-EtG) were purchased from Medichem (Stuttgart,
Germany). Sodium ethyl sulfate was obtained from ABCR
(Karlsruhe, Germany). Deuterated ethyl sulfate (D
5
-EtS) was
prepared by an in-house procedure [3].
Mass spectrometric analyses were performed with a
liquid chromatography-tandem mass spectrometry (LC-MS/
MS) system consisting of an API 365 triple quadrupole
tandem mass spectrometer with a Turbo IonSpray interface
(Applied Biosystems/Sciex, Darmstadt, Germany) and a
HPLC system (three pumps LC-10AD; system controller
SCL-10A Shimadzu, Duisburg, Germany). HPLC sepa-
ration was achieved at 40°C with a polar- endcapped
phenylpropyl reversed phase column (Synergy Polar-RP
250×2 mm, 4 μm) with a guard column (4 mm×2 mm,
same packing m aterial; Phenomenex, Aschaffenburg,
Germany). For isocratic elution, 0.1% formic acid was
used with a flow rate of 0.2 ml/min. Acetonitrile was
added post-column at a flow rate of 0.2 ml via a T union
to enhance the volatility of the eluent. B oth EtG and EtS
were analyzed in one run by a validated MS/MS method
with electrospray ionization (ESI) [14]. The method was
calibrated from 0.45 to 225 μmol/l for EtG and from 0.79
to 397 μmol/l for EtS. The limits of quantitation (LOQ)
in urine ( 0.45 μmol/l for EtG a nd 0.79 μmol/l for EtS)
had been determined during method validation (using
VALISTAT 1.0 software, Arvecon GmbH, Walldorf,
Germany) according to forensic guidelines. For serum,
thesameLOQwasused(0.45μmol/l for EtG and
0.79 μmol/l for EtS), as the s ignal-to-noise ratio for EtG
in serum was 12 at 0.45 μmol/l, and for EtS 10 at
0.79 μmol/l. Intraday precision with serum samples was
11.8% (at 3.28 μm ol/l , n=6) and 8.1% (at 1.90 μmol/ l, n =
7) for EtG, respectively, and 11.0% (at 2.26 μmol/l, n =7)
and 13.3% (at 0.88 μmol/l, n =7) for EtS, respectively.
For EtG, the MS/MS transition with m/z 221/75
(precursor ion/product ion) was used as quantifier, m/z
221/203, 221/113, and 221/85 were used as qualifiers and
m/z 226/75 represented the deuterated standard. The
transitions for EtS were m/z 125/97 (quantifier), 125/80
(qualifier), and 130/98 for the deuterated standard.
Ethanol in urine and serum was determined in duplicate
both by headspace-gas chrom atography with flame ioniza-
tion detector (GC-FID) with tertiary butanol as internal
standard and enzymatically using an alcohol dehydrogenase
method (Cobas Mira S, Roche, Mannheim, Germany with
DRI® Ethyl Alcohol Assay 0037, Microgenics, Passau,
Germany). The serum concentrations have been converted
to blood alcohol concentrations according to forensic
124 Int J Legal Med (2008) 122:123128
guidelines by use of the conversion factor 0.809. Urinary
creatinine was determined by a Roche Hitachi 902 (Roche
Diagnostics, Mannheim, Germany) analyzer using the Jaffé
reaction (DRI® Creati nine -Det ect® Test, Microge nics,
Passau, Germany). Data were processed with Microsoft
Excel 2002. Statistic parameters (correlation coefficient,
significance level) were calculated using SPSS 13.0.1.
Sample preparation
Urine or serum samples (100 μl) were spiked with 30 μlof
a ready-made mixture of D
5
-EtG and D
5
-EtS (containing
0.1 μgD
5
-EtG in 30 μl standard mixture and a constant
amount of D
5
-EtS synthesized by an in-house procedure
[3]), proteins were precipitated from the mixture with
250 μl methanol for urine and 250 μl acetonitrile for serum
samples, respectively. After centrifugation, 270 μl of the
supernatant was evaporated in a vacuum centrifuge and
reconstituted with 140 μl of 0.1% aqueous formic acid
(280 μl for urine samples with a creatinine concentration
higher than 135 mg/dl), and 10 μl aliquots were injected
into the LC-MS/MS system.
Results
The blank blood and urine samples obtained before the start of
drinking were all negative for ethanol, EtG, and EtS. The vol-
unteersconsumedbetween0.50and0.78gethanol/kgbody
mass (see T able 1). The resulting mean peak EtG and mean
peak EtS concentrations in serum were 1.24. 9 μmol/l (mean
2.9±1.3 μmol/l SD, relative SD 43%, median 2.8 μmol/l) and
1.06.4 μmol/l (2.8±1.6 μmol/l, 59%, 2.3 μmol/l), respec-
tively (see Table 2,Fig.1). Peak serum EtG concentrations
were reached between 2.3 and 5.0 h (4.0±0.9 h, 21%, 4.25 h)
after the start of drinking, peak EtS concentrations after 2.1
3.9 h (3.0±0.5 h, 17%, 2.9 h).
Peak blood alcohol concentrations (BAC) were reached
between 1.3 and 2.1 h (1.8±0.2 h, 13%, 1.9 h). Differences
of times for reaching mean peak BAC and mean peak Se
ETS concentrations were between 0.5 and 2.0 h (1.2±0.5 h,
42%, 1.2 h). Differences of times for reaching peak BAC
and peak Se ETG concentrations were between 0.5 and
3.5 h (2.3±0.9 h, 38%, 2.1 h). Differences of times for
reaching peak EtS and peak EtG concentrations in serum
were between 0 and 2.4 h (1.2±0.9 h, 71%, 1.3 h) (see
Table 2).
In seven volunteers, higher molar peak EtS concentra-
tions were found, but in contrast in six persons, the molar
peak EtG concentration was higher than the peak EtS
concentration. Eleven of the volunteers still had EtG in the
last blood sample more than 10 h after the start of drinking,
whereas only two volunteers had an EtS positive last
sample. Remarkably, in the serum samples of one volunteer
(no. 9), no EtS could be detected, although EtG and ethanol
were detectable. In the remaining ten volunteers, the last
EtS-positive serum sample was drawn between 3.8 and
9.5 h after the start of drinking. The molar ratio of peak
EtS/EtG ranged from 0 to 1.31 (0.90±0.35, relative SD
39%, median 1.01). Comparison of peak serum EtG and
EtS c oncentrations from two volunteers indicated, for
volunteer 3, that the peak EtS concentration was 60% of
the EtG concentration, whereas volunteer 13 showed a 1.3
times higher EtS concentration than EtG (see Fig. 2).
Mean peak urinary concentrations of 104805 μmol/l
(401±232 μmol/l, 58%, 409 μmol/l) for EtG and 46
533 μmol/l (266±153 μmol/l, 57%, 283 μmol/l) for EtS
(see Table 2
,Fig.3) were found, which peaked between 5.0
and 7.5 h (6.2±0.9 h, 14%, 6.2 h) (EtG) and 3.1 and 7.4 h
(5.3±1.2 h, 22%, 5.5 h) (EtS). In 12 volunteers, the molar
peak urinary concentration of EtG was higher than that of
a
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
0:00 1:00 2:00 3:00 4:00 5:00 6:00 7:00 8:00 9:00 10:00 11:00 12:00
time [h:min]
BAC [g/ kg]
1
2
3
4
5
6
7
8
9
10
11
12
13
b
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
0:00 1:00 2:00 3:00 4:00 5:00 6:00 7:00 8:00 9:00 10:00 11:00 12:00
time (h:min)
SeEtS [µmol/ L]
c
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
0:00 1:00 2:00 3:00 4:00 5:00 6:00 7:00 8:00 9:00 10:00 11:00 12:00
time (h:min)
SeEtG [µmol/ L]
Fig. 1 Concentrations of ethanol in blood (a), EtS in serum (b), and
EtG in serum (c)(from top to bottom) after consumption of 0.50
0.78 g EtOH/kg body weight
Int J Legal Med (2008) 122:123128 125
EtS. Volunteer 4 showed more urinary EtS than EtG. Four of
the subjects still had EtG in the last urine sample (longer
than 44 h), and the other nine persons had the last EtG-
positive urine sample between 26.6 and 36.1 h after the
start of drinking. EtS showed a slightly different profile, as
one person had an EtS-positive urine in the last sample after
70 h, but this person showed a significant increase in EtS
levels after 42 h, although there was no increase in EtG.
Two persons still excreted EtS in urine after more than
47 h, and from the remaining ten urine samples, EtS had
disappeared between 22.8 and 35.8 h after the start of
drinking. The ratio of molar concentrations of EtS and EtG
ranged from 0.44 to 1.24 (0.70±0.25, 35%, 0.69). Also in
urine, large interindividual differences in peak EtG and EtS
concentrations were observed. The peak EtS concentration
of volunteer 3 was 28% of the peak EtG concentration,
whereas it was 88% of the peak EtG concentration in
volunteer 8.
The following parameters have be en checked for
correlation in pairs: peak concentrations of EtG, EtS, and
ethanol in serum, peak concentrations of EtG, EtS, and
ethanol in urine, areas under the curve (AUC) of EtG, EtS,
and ethanol in serum, c onsume d amount of alcohol,
consumed amount of alcohol per kg body weight, age,
height, body mass, and body mass index (BMI).
Correlations were found for ethanol, EtG, and EtS, when
comparing s erum peak concentrations and AUCs. In
addition, seven highly significant correlations (p 0.001)
were found (see Table 3), using the model of KendallsTau
b correlation. This statistical model was applied, as the
variables were not normally distributed, had a moderate
linear relationship, and because the Kendall Tau b model is
advantageous when outliers occur.
For serum, the AUC of ethanol correlated with the peak
concentration of EtG (correlation coefficient 0.753; signif-
icance level p<0.001) and also with the AUC of EtG
(0.735; p<0.001) and the AUC of EtS (0.857; p<0.001).
Correlations were also found between AUC of EtS and the
peak concentration of EtG (0.805; p<0.001) and also with
the peak concentration of ethanol (0.737; p=0.001), and
with the AUC of EtG (0.761; p<0.001). For urine, there
was a correlation between the peak concentration of EtG
and the peak concentration of EtS (0.684; p=0.001).
In Table 2, the mean peak blood and urine concen-
trations of ethanol, serum and urine EtG and EtS are
compared. As expected, the SDs for ethanol both in urine
and blood were smaller than those for EtG and EtS. One
volunteer even showed no detectable amount of EtS in
serum, whereas ethanol and EtG wer e found in blood and
EtS and EtG in urine as well. Peak ETG concentration in
this volunteers serum was 40% of the average value of all
a
0
0.1
0.2
0.3
0.4
0.5
0.6
0:00 2:24 4:48 7:12 9:36 12:00
time (h:min)
BAC [g ‰]
0.00
1.00
2.00
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
c EtG and c EtS [µmol/ L]
Vol 3 SeEtOH
Vol 3 SeEtS
Vol 3 SeEtG
b
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
0:00 1:12 2:24 3:36 4:48 6:00 7:12 8:24 9:36 10:48
time (h:mm)
BAC [g ‰]
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
c EtG and c EtS [
µ
mol/ L]
Vol 13 SeEtOH
Vol 13 SeEtG
Vol 13 SeEtS
Fig. 2 a, b Blood ethanol (BAC), serum EtS and EtG in two
volunteers. a volunteer 3 (0.52 g EtOH/kg body weight), b volunteer
13 (0.78 g EtOH/kg body weight)
a
0
100
200
300
400
500
600
700
800
900
0:00 12:00 24:00 36:00 48:00 60:00 72:00
time (h:min)
EtG [µmol/ L]
1
2
3
4
5
6
7
8
9
10
11
12
13
b
0
2
4
6
8
18:00 30:00 42:00 54:00 66:00
0
100
200
300
400
500
600
0:00 12:00 24:00 36:00 48:00 60:00 72:00
time (h:min)
EtS [µmol/ L]
1
2
3
4
5
6
7
8
9
10
11
12
13
0
2
4
6
8
10
18:00 30:00 42:00 54:00 66:00
Fig. 3 Concentrations of EtG (a)andEtS(b)inurineafter
consumption of 0.500.78 g EtOH/kg body weight
126 Int J Legal Med (2008) 122:123128
volunteers, peak EtS concentration in urine 26%, and peak
EtG concentration in urine 18% of the average value,
respectively.
Discussion
EtS and EtG were detectable in urine and serum for a
longer period of time than ethanol itself. In serum, EtS was
detectable for about twice as long as ethanol, and EtG, for
even longer. In urine, EtG was detectable for up to 10 times
longer than ethanol, EtS, 38 times longer. There were
large variations in the time differences for reaching
maximum concent rations of blood alcohol, serum EtG,
and serum EtS. Some correlations could be found, but most
of them involved at least one AUC, which is not very
useful in spot sample applications. The only fully non-AUC
correlation occurred between the maximum concentrations
of EtG and EtS in urine.
In former studies, it has been shown that glucuronyl-
transferases and sulfotransferases show polymorphism [5,
13], and furthermore, induction of the glucuronyltransferase
UGT1A1 by alcohol has been reported [8]. By inclusion of
enzymatic formation, distributio n, and elimination, a kinetic
model for EtG was recently postulated by Droenner et al.
[4], but none has been calculated for EtS, yet. However, our
study clearly shows that there are huge interindividual
differences particularly in sulfoconjugation. In addition to
polymorphisms of conjugating enzymes, differences in their
activity, or expression, the sulfation might be influenced by
other factors, such as nutrition as proposed in the twin
study of Nash et al. [9]. Therefore, and due to the relatively
low number of individuals tested, we could not describe a
new kinetic model for EtS applicable for all volunteers.
Furthermore, we detected an unexpected incre ase in urinary
EtS after 42 h in volunteer 13 who had shown normal
formation and urinary excretion of both EtG and EtS in
parallel in several preceding drinking studies. This person
was compliant concerning abstinence from ethanol and an
incidental uptake of alcohol leading to 94 μmol/l EtG in
urine would also have rais ed the EtG levels, which was not
observed, and therefore, can be excluded. The reason for
this increase has to be found elsewhere, e.g., uptake of EtS
with food or nonalcoholic beverages. To prove this, further
studies on this aspect have to be performed. To collect
information for further evaluation of both markers, we
included both markers in our routine analysis method,
which is used for monitoring compliance of patients in
ethanol withdrawal therapy and for forensic purposes.
Testing for EtG and EtS in parallelwhich is not more
time-consumi ng than EtG aloneis recommended for
forensic cases, as excretion in urine is qualitatively very
similar, although interindividual variances do occur.
Furthermore, stability issues, due to bacterial contamination
caused by e.g., urinary tract infections have been raised,
which resulted in a degradation of EtG but not EtS [7].
Acknowledgements The authors wish to thank the Bund gegen
Alkohol und Drogen im Straßenverkehr e.V. for the funding of the
study.
References
1. Barbour AD (2001) Simplified estimation of Widmark r values
by the method of Forrest. Sci Justice 41:5354
2. UGT Alleles Nomenclature Home Page. UGT Nomenclature
Committee. June 2005. [21.03.2007]. http://galien.pha.ulaval.ca/
alleles/alleles.html
3. Dresen S, Weinmann W, Wurst FM (2004) Forensic confirmatory
analysis of ethyl sulfatea new marker for alcohol consumption
by liquid-chromatograp hy/electrospray ionization/tandem mass
spectrometry. J Am Soc Mass Spectrom 15:16441648
4. Droenn er P, Schmitt G, Aderjan R, Zimmer H (2002) A kinetic
model describing the pharmacokinetics of ethyl glucuronide in
humans. Forensic Sci Int 126:2429
5. Foti RS, Fisher MB (2005) Assessment of UDP-glucuronosyl-
transferase catalyzed formation of ethyl glucuronide in human
liver microsomes and recombinant UGTs. Forensic Sci Int
153:109116
6. Helander A, Beck O (2004) Mass spectrometric identification of
ethyl sulfate as an ethanol metabolite in humans. Clin Chem
50:936937
7. Helander A, Dahl H (2005) Urinary tract infection: a risk factor
for false-negative urinary ethyl glucuronide but not ethyl sulfate in
the detection of recent alcohol consumption. Clin Chem 51:1728
1730
8. Kardon T, Coffey MJ, Banhegyi G, Conley AA, Burchell B,
Mandl J, Braun L (2000) Transcriptional induction of bilirubin
UDP-glucuronosyltransrase by ethanol in rat liver. Alcohol
21:251257
9. Nash RM, Stein L, Penno MB, Passananti GT, Vesell ES (1984)
Sources of interindividual variations in acetaminophen and
antipyrine metabolism. Clin Pharmacol Ther 36:417430
10. Politi L, Morini L, Groppi A, Poloni V, Pozzi F, Polettini A (2005)
Direct determination of the ethanol metabolites ethyl glucuronide
and ethyl sulfate in urine by liquid chromatography/electrospray
tandem mass spectrometry. Rapid Comm Mass Spectrom
19:13211331
11. Schloegl H, Dresen S, Spaczynski K, Stoertzel M, Wurst FM,
Weinmann W (2006) Stability of ethyl glucuronide in urine, post-
mortem tissue and blood samples. Int J Legal Med 120:8388
12. Schneider H, Glatt H (2004) Sulpho-conjugation of ethanol in
humans in vivo and by individual sulphotransferase forms in vitro.
Biochem J 383:543549
13. Thomae BA, Rifki OF, Theobald MA, Eckloff BW, Wieben ED,
Weinshilboum RM (2003) Human catecholamine sulfotransferase
(SULT1A3) pharmacogenetics: functional genetic polymorphism.
J Neurochem 87:809819
14. Weinmann W, Schaefer P, Thierauf A, Schreiber A, Wurst FM
(2004) Confirmatory analysis of ethylglucuronide in urine by
liquid-chromatography/electrospray ionization/tandem mass spec-
trometry according to forens ic guidelines. J Am Soc Mass
Spectrom 15:188193
Int J Legal Med (2008) 122:123128 127
15. Wurst FM, Metzger J (2002) The ethanol conjugate ethyl
glucuronide is a useful marker of recent alcohol consumption.
Alcohol Clin Exp Res 26:11141119
16. Wurst FM, Skipper GE, Weinmann W (2003) Ethyl glucuronide
the direct ethanol metabolite on the threshold from science to
routine use. Addiction 98(Suppl 2):5161
17. Wurst FM, Wiesbeck GA, Metzger JW, Weinmann W (2004) On
sensitivity, specificity, and the influence of various parameters on
ethyl glucuronide levels in urineresults from the WHO/ISBRA
study. Alcohol Clin Exp Res 28:12201228
18. Wurst FM, Alling C, Aradottir S et al (2005) Emerging biomarkers: new
directions and clinical applications. Alcohol Clin Exp Res 29:46547 3
128 Int J Legal Med (2008) 122:123128
    • "After the consumption of 0.50-0.78 g ethanol/kg body weight, EtG and EtS remained detectable in blood for up to more than 10 h (with a detection window twice the one for ethanol) [53]. While multiple methods for the quantification of EtG and EtS in blood have been published [6,12,[54][55][56][57][58][59][60][61][62], there is currently only one report on their quantification in DBS [63]. "
    [Show abstract] [Hide abstract] ABSTRACT: Monitoring of alcohol consumption by living persons takes place in various contexts, amongst which workplace drug testing, driving under the influence of alcohol, driving licence regranting programs, alcohol withdrawal treatment, diagnosis of acute intoxication or fetal alcohol ingestion. The matrices that are mostly used today include blood, breath and urine. The aim of this review is to present alternative sampling strategies that allow monitoring of the alcohol consumption in living subjects. Ethanol itself, indirect (carbohydrate deficient transferrin, CDT%) as well as direct biomarkers (ethyl glucuronide, EtG; ethyl sulphate, EtS; fatty acid ethyl esters, FAEEs and phosphatidylethanol species, PEths) of ethanol consumption will be considered. This review covers dried blood spots (CDT%, EtG/EtS, PEths), dried urine spots (EtG/EtS), sweat and skin surface lipids (ethanol, EtG, FAEEs), oral fluid (ethanol, EtG), exhaled breath (PEths), hair (EtG, FAEEs), nail (EtG), meconium (EtG/EtS, FAEEs), umbilical cord and placenta (EtG/EtS and PEth 16:0/18:1). Main results, issues and considerations specific to each matrix are reported. Details about sample preparation and analytical methods are not within the scope of this review.
    Article · May 2016
    • "EtS has a molecular weight of 126 g/mol, and represents, like EtG, a secondary elimination pathway for alcohol. EtS is detectable in varying interindividual concentrations (Dresen et al., 2004; Halter et al., 2008; Helander and Beck, 2004; Wurst et al., 2006). An immunochemical detection test is currently not commercially available for EtS. "
    [Show abstract] [Hide abstract] ABSTRACT: Background: Alcohol-related disorders are common, expensive in their course, and often underdiagnosed. To facilitate early diagnosis and therapy of alcohol-related disorders and to prevent later complications, questionnaires and biomarkers are useful. Methods: Indirect state markers like gamma-glutamyl-transpeptidase, mean corpuscular volume, and carbohydrate deficient transferrin are influenced by age, gender, various substances, and nonalcohol-related illnesses, and do not cover the entire timeline for alcohol consumption. Ethanol (EtOH) metabolites, such as ethyl glucuronide, ethyl sulfate, phosphatidylethanol, and fatty acid ethyl esters have gained enormous interest in the last decades as they are detectable after EtOH intake. Results: For each biomarker, pharmacological characteristics, detection methods in different body tissues, sensitivity/specificity values, cutoff values, time frames of detection, and general limitations are presented. Another focus of the review is the use of the markers in special clinical and forensic samples. Conclusions: Depending on the biological material used for analysis, ethanol metabolites can be applied in different settings such as assessment of alcohol intake, screening, prevention, diagnosis, and therapy of alcohol use disorders.
    Full-text · Article · Sep 2015
    • "The differences in gender are almost negligible in the age group ranging from 18-24 years [2]. The identification and quantification of biomarker, specific for the consumption and/or abuse of alcohol, is of relevance for forensic purposes since it offers the possibility of distinguishing between the intake of alcohol in matrices such as blood and serum (in the short term), urine (in the medium term) and hair and meconium (in the long term) [3][4][5]. Ethyl glucuronide (EtG), being a direct metabolite of "
    [Show abstract] [Hide abstract] ABSTRACT: The determination of ethyl glucuronide (EtG), a stable and sensitive marker that is specific to alcohol intake, finds many applications both in the forensic toxicology and clinical fields. The aim of the study is to examine the possibility of using a cadaveric biological matrix, vitreous humor (VH), to determine EtG as a marker of recent ethanol use. The blood, taken from the femoral vein, and the VH were obtained from 63 autopsy cases. Analysis of the EtG was performed using an LC/MS/MS system. Analyses of the ethanol and putrefaction biomarkers, such as acetaldehyde and n-propanol, were performed using the HS-GC/FID technique in both the matrices. In 17 cases, both ethanol and EtG were absent in both matrices.Nineteen cases presented ethanol in blood from 0.05 to 0.30 g/L, EtG-Blood concentration from 0.02 to 3.27 mg/L, and EtG-VH concentration from 0.01 mg/L to 2.88 mg/L. Thirteen cases presented ethanol in blood > 0.05 g/L but EtG concentration in blood and VH lower than 0.01 mg/L, are part of these 8 samples presented acetic aldehyde and n- propanol in blood or VH, means identification of putrefaction indicators. Fourteen cases presented ethanol in blood > 0.46 and EtG concentration in blood and VH higher than 0.01 mg/L. The determination of EtG in biological material is important in those cases where the intake of ethanol appears doubtful, as it allows us to exclude the possibility of any post-mortem formation of ethanol.
    Article · Mar 2015
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