RESUMO
BACKGROUND: Ethyl glucuronide (EtG) in urine is considered a specific marker of recent ethanol consumption. There is an ongoing debate about whether inhalation or transdermal resorption of sanitizer ethanol is the underlying cause for positive EtG findings after hand disinfection. METHODS: Desderman(®) pure (Schülke & Mayr GmbH, Norderstedt) with 78.2g 96% (v/v) ethanol/100g and approx. 10% 2-propanol was used for multiple hand disinfection without and under an exhauster. Simulating a common working day in a clinic, 5 co-workers of our lab used the sanitizer 32 fold within 8h and 2 persons were merely exposed to the sanitizer vapor but without any dermal sanitizer contact. Any additional ethanol intake or exposition was reliably excluded. Spot urine was collected at baseline, after 1, 2, 4, 6 14, and finally 24h after the first sanitizer use. A validated LC-MS/MS was used for MRM and MS(3) of EtG and qualitative analyses of ethyl sulfate and 2-propyl glucuronide. RESULTS: Multiple hand disinfection caused positive EtG findings of up to 2.1mg/L or 1.7mg/g creatinine in 4 out of 5 test persons and even of 0.6mg/L or 0.8mg/g for 2 controls which were merely exposed to the sanitizer vapor but without any sanitizer contact. EtG results between the clinical (0.5mg/g) and the forensic (0.1mg/g) cut-off were obtained even 6h after the last sanitizer exposition. An exhauster prevented the sanitizer vapor inhalation and reduced the EtG excretion to mostly below the detection limit of 0.02mg/g. The maximum value was 0.09mg/g. Ethyl sulfate and 2-propyl glucuronide (2-PpG) were detectable only in the EtG positive samples. 2-PpG is a metabolite of 2-propanol, which is quite frequently used in disinfectants. Thus, the detection of this substance can be used in cases of odd EtG results as an indicator of (unintended) sanitizer exposition. CONCLUSION: Ethanol from hand sanitizers is predominantly incorporated by the respiratory tract but not via the skin. It can cause a distinct ethyl glucuronide excretion and thus analytically true-positive but forensically false-positive EtG findings in the urine of ethanol abstaining persons. Since accidental ethanol inhalation can occur quite frequently in the working place or even private household, such a situation should always be considered when EtG is used as a marker of recent ethanol consumption.
Assuntos
Glucuronatos/urina , Higienizadores de Mão/química , Volatilização , 2-Propanol/urina , Anti-Infecciosos Locais/análise , Cromatografia Líquida , Etanol/análise , Humanos , Espectrometria de Massas , Ésteres do Ácido Sulfúrico/urinaRESUMO
AIM: A drug and alcohol withdrawal rehabilitation centre requested an analysis for "Krypton" in urine of a former opiate-addictive woman. She showed an altered clinical picture and behaviour with miosis, itchiness, agitation, and moderate euphoria after 3 months of until than successful treatment. Literature search revealed that "Krypton" is said to contain "Kratom" (leaves of Mitragyna speciosa), but could also contain O-desmethyltramadol (European Monitoring Centre for Drugs and Drug Addiction thematic paper "Spice"). METHODS: Immunological drug screenings were done with test strips (nal von minden, Regensburg, Germany) and with cloned enzyme donor immunoassay (Microgenics, Passau, Germany). "Kratom" alkaloids and tramadol (metabolites) were analyzed by LC-MS/MS (ThermoFisher Scientific Quantum Ultra Triple Quadrupole mass spectrometer). RESULTS: Immunoassays were negative for amphetamines, barbiturates, benzodiazepines, benzoylecgonine, buprenorphine, ethylglucuronide, methadone (metabolite), opiates, oxycodone, and THC-COOH, and test strips were negative for tramadol and its metabolites (cut-off 10 mg/L for O-desmethyltramadol). LC-MS/MS detected the "Kratom" alkaloids mitragynine, speciociliatine, speciogynine, mitraciliatine, and paynantheine and approximately 9mg/L O-desmethyltramadol, but no tramadol and N-desmethyltramadol. DISCUSSION: The detection of M. speciosa alkaloids is a proof of "Kratom" abuse. Confronted with the analysis data, the patient admitted to have consumed 3-4 infusions of "Krypton". The origin of the O-desmethyltramadol is unclear. Tramadol abuse is unlikely since tramadol and N-desmethyltramadol (physiologically occurring in urine after tramadol intake) were not detectable. Consumption of a "Krypton" product spiked with O-desmethyltramadol could explain our findings and the patient's clinical picture. This would be in agreement with a most recent report about spiking apparently natural herbal mixtures with the synthetic opioid O-desmethyltramadol. CONCLUSION: Analysis of "Kratom" abuse should not be restricted to M. speciosa alkaloids, but should also consider synthetic drugs which could be added to the herbal mixtures. Mass spectrometry based drug screenings will gain importance to keep pace with the dynamic drug market.
Assuntos
Alcaloides/urina , Mitragyna/química , Extratos Vegetais/urina , Folhas de Planta/química , Tramadol/análogos & derivados , Adulto , Feminino , Toxicologia Forense , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Alcaloides de Triptamina e Secologanina/urina , Detecção do Abuso de Substâncias , Transtornos Relacionados ao Uso de Substâncias/diagnóstico , Tramadol/urinaRESUMO
BACKGROUND: Urine-ethyl glucuronide (EtG) concentrations are considered as a specific marker of recent alcohol consumption. We describe false-positive EtG screening results by the DRI ethyl glucuronide enzyme immunoassay caused by chloral hydrate intake. METHODS: Urine-EtG-screening: DRI EtG enzyme immunoassay (Thermo Fisher Scientific Microgenics) on a Hitachi 912 analyzer. EtG- and ethyl sulfate (EtS) confirmatory analysis: LC-MS/MS with an ESI source in the negative ionization, selective reaction monitoring mode. PATIENT: ethanol-abstaining women under buprenorphine-treatment (medication with levetiracetam, gabapentin, clomethiazol and chloral hydrate). Proband: self-medication with 500 mg chloral hydrate after a 5-day ethanol abstinence. EtG analysis for both in subsequent urines. Check for cross reactions of the pharmaceuticals with the EtG immunoassay by addition of pure substance (2 g/L each) to EtG-free urine. RESULTS: EtG concentrations up to 8.0 mg/L or 7.0 mg/g creatinine (cut-off 0.5 mg/L or mg/g) for the patient and up to 0.28 mg/L or 0.35 mg/g for the control subject (after 500 mg chloral hydrate) were obtained by the immunoassay. LC-MS/MS could not confirm these EtG results. In fact, EtG and/or EtS were not detectable in any of the urine samples by LC-MS/MS (lower limit of detection 0.01 mg/L). Cross reactions of the pharmaceuticals, incl. the chloral hydrate metabolites trichloroethanol and trichloroacetic acid, with the DRI EtG immunoassay results were ruled out (by spiking experiments) as the underlying cause for the false-positive EtG immunoassay results. CONCLUSIONS: Trichloroethyl glucuronide as an important chloral hydrate metabolite remains the most probable cross reacting substance with the DRI EtG immunoassay (unproven because of lack in pure standard). The chloral hydrate self-medication experiment clearly points to an association of the false-positive EtG immunoassay results and chloral hydrate intake. Chloral hydrate medication has to be considered as a cause for false-positive EtG screening results by the DRI EtG immunoassay even in cases with regular chloral hydrate treatment (250-1000 mg) and the more in patients with chloral hydrate tolerance (taking g/day). It is recommended that positive EtG immunoassay results always be confirmed by a more specific technique such as LC-MS/MS, including ethyl sulfate as a second minor ethanol metabolite.