背景:尿液中的乙基葡糖苷酸(EtG)被认为是最近乙醇消耗或乙醇暴露的标志。叔丁醇主要用作溶剂和中间化学品。像叔戊醇一样,叔丁醇在互联网论坛上讨论作为乙醇替代。我们讨论了通过排泄不同的醇葡糖苷酸(EtG同源物)进行的假阳性免疫EtG筛查,多甲托克科曼病患者尿液中主要是叔丁基葡糖苷酸。
方法:在BeckmanCoulterAU680分析仪上通过DRIEtG酶免疫测定(ThermoFisherScientificMicrogenics)分析了长期滥用多种物质(包括溶剂)的住院患者的三个连续尿液样本,EtG的内部LC-MS/MS,1-丙基,2-丙基,1-丁基,2-丁基,和叔丁基葡糖苷酸,和自由同类物质甲醇的内部顶空GC-FID,1-丙醇,2-丁酮,2-丁醇,异丁醇,1-丁醇,3-甲基-1-丁醇,2-甲基-1-丁醇,另外对于乙醇,丙酮,2-丙醇,叔丁醇和2-甲基-2-丁醇。
结果:EtG免疫测定产生两个阳性尿液样本(0.2和0.6mg/L或0.1和0.2mg/g肌酐;截止值0.1mg/L),经LC-MS/MS检测EtG阴性(截止值0.1mg/L),但对叔丁基葡糖苷酸(3.7和27.1mg/L)呈阳性,2-丁基葡糖苷酸(1.1和3.5mg/L),和2-丙基葡糖苷酸(0.1和0.4mg/L)。顶空GC-FID检测叔丁醇(0.97和4.01mg/L),甲醇(0.96和0.62mg/L),2-丁酮(0.84和1.65mg/L),和2-丁醇(0.04和0.09mg/L),但没有乙醇和2-甲基-2-丁醇.
结论:EtG同系物的交叉反应,怀疑叔丁醇或异丁烷滥用后,主要是叔丁基葡糖苷酸,解释了EtG免疫测定结果的假阳性。未来的研究可以解决尿液中的醇葡糖醛酸苷(EtG同系物)作为(a)对烷烃或其相应的醇代谢产物的生物标志物,以及(b)作为使用“老”-众所周知的醇(如叔丁醇或叔戊醇)的标记容易获得,便宜,有效和“不可检测”的乙醇替代品或“新”的精神活性醇。
BACKGROUND: Ethyl glucuronide (EtG) in urine is considered a marker of recent ethanol consumption or ethanol exposition. tert-Butanol is primarily used as a solvent and intermediate chemical. Like tert-amyl alcohol, tert-butanol is discussed in Internet forums as ethanol replacement. We discuss false-positive immunological EtG screenings by excretion of different alcohol glucuronides (EtG homologs), mainly tert-butyl glucuronide in urine of a polytoxikomanic in-patient.
METHODS: Three consecutive urine samples from an in-patient with a long history of multiple substance abuse including solvents were analyzed by DRI EtG enzyme immunoassay (ThermoFisher Scientific Microgenics) on a Beckman Coulter AU680 analyzer, an in-house LC-MS/MS for EtG, 1-propyl, 2-propyl, 1-butyl, 2-butyl, and tert-butyl glucuronide, and an in-house headspace GC-FID of free congener substances methanol, 1-propanol, 2-butanone, 2-butanol, isobutanol, 1-butanol, 3-methyl-1-butanol, 2-methyl-1-butanol, and additionally for ethanol, acetone, 2-propanol, tert-butanol and 2-methyl-2-butanol.
RESULTS: EtG immunoassay yielded two positive urine samples (0.2 and 0.6mg/L or 0.1 and 0.2mg/g creatinine; cut-off 0.1mg/L) which were tested EtG negative by LC-MS/MS (cut-off 0.1mg/L) but positive for tert-butyl glucuronide (3.7 and 27.1mg/L), 2-butyl glucuronide (1.1 and 3.5mg/L), and 2-propyl glucuronide (0.1 and 0.4mg/L). Headspace GC-FID detected tert-butanol (0.97 and 4.01mg/L), methanol (0.96 and 0.62mg/L), 2-butanone (0.84 and 1.65mg/L), and 2-butanol (0.04 and 0.09mg/L), but no ethanol and no 2-methyl-2-butanol.
CONCLUSIONS: Cross-reaction of EtG homologs, mainly tert-butyl glucuronide after suspected tert-butanol or isobutane abuse, explains the false-positive EtG immunoassay findings. Future investigations could address the usefulness of alcohol glucuronides (EtG homologs) in urine as (a) biomarkers of an exposition to alkans or their corresponding alcohol metabolites and (b) as markers for using \"old\"-well known alcohols like tert-butanol or tert-amyl alcohol as easy to obtain, cheap, potent and \"undetectable\" ethanol replacements or \"New\" Psychoactive Alcohols.