Postcollection synthesis of ethyl glucuronide by bacteria in urine may cause false identification of alcohol consumption.
BACKGROUND
Ethyl glucuronide (EtG) is a minor ethanol metabolite used as a specific marker to document recent alcohol consumption; confirm abstinence in treatment programs, workplaces, and schools; and provide legal proof of drinking. This study examined if bacterial pathogens in urine may enable postsampling synthesis of EtG and ethyl sulfate (EtS) from ethanol, leading to clinical false-positive results.


METHODS
Urine specimens with confirmed growth of Escherichia coli, Klebsiella pneumoniae, or Enterobacter cloacae were stored at room temperature in the presence of ethanol. Ethanol was either added to the samples or generated by inoculation with the fermenting yeast species Candida albicans and glucose as substrate. EtG and EtS were measured by LC-MS.


RESULTS
High concentrations of EtG (24-h range 0.5-17.6 mg/L) were produced during storage in 35% of E. coli-infected urines containing ethanol. In some specimens that were initially EtG positive because of recent alcohol consumption, EtG was also sensitive to degradation by bacterial hydrolysis. In contrast, EtS was completely stable under these conditions.


CONCLUSIONS
The presence of EtG in urine is not a unique indicator of recent drinking, but might originate from postcollection synthesis if specimens are infected with E. coli and contain ethanol. Given the associated risks for false identification of alcohol consumption and false-negative EtG results due to bacterial degradation, we recommend that measurement of EtG be combined with EtS, or in the future possibly replaced by EtS.
Helander
Anders
A
Department of Clinical Neuroscience, Karolinska Institute, Karolinska University Hospital, Stockholm, Sweden. anders.helander@ki.se
Olsson
Ingrid
I
Dahl
Helen
H
D000428:Alcohol Drinking / Q000652:urine*; D016972:Enterobacter cloacae / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D000431:Ethanol / Q000652:urine*; D005189:False Positive Reactions; D005965:Glucuronates / Q000096:biosynthesis / Q000652:urine*; D006801:Humans; D006868:Hydrolysis; D007711:Klebsiella pneumoniae / Q000378:metabolism*; D013048:Specimen Handling; D015813:Substance Abuse Detection / Q000379:methods*
D005965:Glucuronates; C093924:ethyl glucuronide; D000431:Ethanol