Tobacco and alcohol use during pregnancy are serious public health concerns and result in adverse developmental outcomes. Identifying in utero exposure is often achieved through meconium analysis or via maternal self-report. In this study, we analyzed fetal liver and placenta to examine second trimester alcohol and smoking exposure. Methods A validated liquid chromatography-tandem mass spectrometry method for simultaneous analysis of nicotine and its metabolites and alcohol markers (ethyl glucuronide: EtG and ethyl sulfate: EtS) was employed to analyze 193 fetal liver and 48 placenta (n=47 paired) samples from electively-terminated pregnancies. Results EtG, EtS and nicotine markers' limits of detection were 0.7-20 ng/g in fetal samples. Ninety-eight fetal liver and 23 placenta samples were EtG/EtS-positive, while 137 liver and 25 placenta samples were positive for tobacco exposure. When both alcohol markers were present in samples, EtG/EtS ratios were 1.6-11.1 in 17 livers and 2.5-31.1 in 10 placentas. Median [range] summed tobacco marker concentrations were 422 [1.0-2776] and 154 [1.6-1621] ng/g in livers and placentas. Median EtG and nicotine marker concentrations were higher in liver than placenta in paired samples. Strong evidence of exposure occurred in 11 and 22 pairs, respectively, when both samples were positive for alcohol and/or tobacco markers. Conclusions These paired fetal liver and placenta alcohol and tobacco data provided a unique means for examining the effects of in utero exposure, a critical first step in selecting fetal samples for proteomic and RNA-sequencing studies that could provide mechanisms for adverse developmental outcomes.
Although kombucha is a popular fermented beverage, the presence of alcohol markers has not been well studied despite being potential indicators of unintentional impairment. Ethyl glucuronide (EtG) and ethyl sulfate (EtS) were measured in oral fluid and urine collected after consumption of regular or hard kombucha. Participants drank within 20 min and provided all urine voids for 12 h, the first urine voids on days 2 and 3, and oral fluid specimens at fixed time points for 48 h. Screening employed liquid chromatography–tandem mass spectrometry (LC–MS-MS; EtS, 25 ng/mL cutoff [oral]; 100 ng/mL cutoff [urine]; EtG, 500 ng/mL cutoff [urine] and immunoassay (IA; EtG, 500 ng/mL cutoff[urine]). After consuming regular kombucha (n = 12 participants), EtS was not detected in oral fluid but both markers were detected by LC–MS-MS in urine specimens within the first 5 voids from 83% of participants with median (range) concentrations of 240 (100–3,700) ng/mL for EtS and 830 (530–2,200) ng/mL for EtG. Neither marker was positive by IA nor LC–MS-MS after day 1. After consuming hard kombucha (n = 7 participants), 2 (2.8%) of the 70 collected oral fluid specimens tested positive for EtS 3 h after consumption; however, 21 (30%) had EtS levels above the limit of detection (LOD, 10 ng/mL) after 0.5–8 h. Both markers were detected in urine specimens from all participants with median (range) concentrations of 3,381 (559–70,250) ng/mL for EtS and 763 (104–12,864) ng/mL For EtG. Urine specimens were negative for EtG and EtS by the end of the 48-hour study.
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