乙基葡萄糖醛酸苷的检测及其在法医毒理学中的应用

乙基葡萄糖醛酸苷（ethyl glucuronide,EtG）是乙醇在人体内的特异性代谢物。迄今,已有相当数量的文献针对EtG的检测方法、代谢动力学及实际应用等进行了深入探讨。本文综述了EtG的各种检测方法、EtG含量与乙醇摄入量的相互关系以及如何利用EtG含量区分是否酗酒、判断尸体中乙醇来源等,阐述EtG在法医毒理学领域的重要意义。     

唾液中乙醇含量检测试剂条的研究

目的根据唾液和血液中乙醇含量相关性的实验结果,建立了一种简便快速、准确可靠的检测唾液中乙醇含量的方法。方法本方法利用酶学原理,将一定量乙醇氧化酶(ALO)和过氧化物酶以及底物四甲基联苯胺(TMB)固定于试剂条上,当样本中含有乙醇时,酶学反应使底物TMB显色,通过比对反应的不同颜色,对样本中乙醇质量浓度进行半定量。结果用本方法检测300个自愿者的唾液,和用GC/MS法对照检测志愿者唾液中的乙醇含量,定量结果基本一致。本产品检测过程仅需2min,其检测的阈值为0.1mg/mL,敏感度为96.5%,特异性为91%,准确性为94.7%。结论采用酶学方法制备的乙醇含量检测试剂条,通过显色反应对唾液中的乙醇含量进行半定量检测,其特点为快速简便、准确可靠,适合现场使用。     

两种不同方法同步检测血清中乙醇浓度的对比分析

目的对同一临床实验室检测血液中乙醇含量的不同检测系统进行评估,探讨不同检测系统中乙醇的测定结果是否具有可比性,为临床实验室认可提供参考依据。方法收集2012年7月至2012年12月来我院进行交通事故乙醇鉴定的样品120例。分别用微粒子酶免化学发光法和顶空气相色谱法检测血液中乙醇的含量。用SPSS16.0软件统计处理。结果两种方法间分析具有良好的可比性和相关性,差异没有显著性意义。结论应重视分析不同方法不同仪器间结果的比对工作,确保检验结果的准确性和一致性。     

人体内乙醇含量的检测及结果分析述评

过量饮酒已被列为世界公共卫生的主要问题之一,近4%的死亡与酒精相关[1]。酒精会刺激人的神经系统,影响人的正常行为。酒后驾车和酒后滋事已成为影响交通安全和社会和谐的一个重要因素。因此,人体内乙醇含量的检测在交通安全和法庭科学领域具有重要的意义。本文通过文献调研,对乙醇检测的样本采集与保存、检测方法及结果等进行了分析,希望能为从事乙醇检测鉴定的工作人员提供参考和帮助。     

加热时间对血醇含量检验的影响

本文通过对标准品不同加热平衡时间建立校准曲线,来计算已知不同浓度级别血样中的乙醇含量,通过计算相对相差,来比较加热平衡时间对血液乙醇含量检验结果的影响。通过试验方法可以得出,血液乙醇含量检验的影响因素较多,比如检验方法、顶空瓶的大小、色谱拄温度、加热平衡时间、血样标准品的制备等等,但是单从加热平衡时间来看,对血液乙醇含量的检验结果有一定的影响,经过试验发现顶空瓶平衡加热9分钟,是较为快速、对检验结果影响较小的检验条件,这为基层检测单位科学、准确、快速检验涉嫌酒驾人员血液样本中乙醇含量,提供了可靠、准确的科学依据。     

采血管中添加剂对血样中乙醇含量的影响

目的：探讨不同种类采血管对血样中乙醇含量检测结果的影响。方法分别用7种一次性真空采血管[无抗凝剂管、促凝剂管、分离胶-促凝剂管、枸橼酸钠（1∶4）管、枸橼酸钠（1∶9）管、柠檬酸钠（9∶1）管、EDTA-K2管]采集10名志愿者饮酒后2 h血液,用顶空气相色谱法检测血样中乙醇含量。结果相同血样用不同的采血管,其乙醇含量检测结果不同,依次为分离胶-促凝剂管〉促凝剂管〉无抗凝剂管〉EDTA-K2管〉枸橼酸钠（1∶9）管〉枸橼酸钠（1∶4）管,柠檬酸钠（9∶1）管与枸橼酸钠（1∶9）管检测结果基本一致。结论采集涉嫌酒后驾驶的驾驶员血样,应选用一次性真空采血管,首选EDTA-K2管。     

血液样品中乙醇稳定性的实验研究

目的考察在不同存放条件下含乙醇的血液样品中乙醇浓度的变化情况。方法采用顶空-气相色谱法,以异丙醇为内标,对存放条件不同的血液样品中乙醇进行检测。结果冷冻(-10℃)条件存放1至30天,血液样品中乙醇含量无显著变化;冷藏(4℃)条件存放1至30天,血液样品中乙醇含量变化不显著;室温(28℃~33℃)条件存放1至30天,血液样品中乙醇含量显著改变。结论含有乙醇的血液样品,在冷冻、冷藏条件下可较稳定的存放30天;在室温条件下存放30天血液样品中乙醇浓度发生显著变化,不可在此条件下存放。     

血液中乙醇检测结果的法医学分析

目的对交通事故中血液中乙醇检测结果进行法医学分析。方法从检测方法、血液采集方法、采集时间、血液保存、尸体腐败、饮酒量与血液中乙醇质量浓度关系等方面进行血液中乙醇检测结果的法医学分析。结果检测方法、血液采集方法、采集时间、血液保存、尸体腐败等因素直接影响血液中乙醇检测结果。结论为保证交通执法的公正性，对血液中乙醇检测结果应当作法医学分析。     

HS-GC内标曲线法测定血中乙醇含量的不确定度评估

目的建立自动顶空-气相色谱(HS-GC)内标曲线法测定血中乙醇含量的不确定评估方法。方法从分析测定程序着手,依据不确定度评定的指导性文件,分析不确定度来源,量化不确定度分量,计算检测结果的合成标准不确定度和扩展不确定度。结果各相对不确定度来自于检材重复性检测为3.4%,乙醇标准溶液为0.71%,检材为0.61%,叔丁醇内标溶液为0.41%,标准曲线为1.1%,气相色谱仪为1.3%,血液中乙醇的相对扩展不确定度为3.9%。结论血液中乙醇含量的不确定度主要来源于检材重复性检测、气相色谱仪、乙醇标准曲线。     

酶促试纸条测定唾液中乙醇含量的研究

生化酶促反应使试纸与含有乙醇的液体接触后显色,根据颜色的不同确定被测液体中的乙醇含量,依此研制出酶促试纸条。通过志愿者实验,同时收集血液和唾液, 比较酶促试纸条测定唾液中乙醇含量与顶空气相色谱检测血醇和唾醇的结果。考察酶促试纸条的影响因素。通过提高缓冲液浓度至0.6mol/L和减少乙醇氧化酶的量改进酶促试纸条,使相同乙醇浓度的酸碱两种唾液间的差异显著缩小。所研制的酶促试纸条具有成本低、操作简便、快速的优点。     

Testing Antemortem Blood for Ethanol Concentration from a Blood Kit in a Refrigerator Fire

The stability of ethanol in antemortem blood stored under various conditions has been widely studied. Antemortem blood samples stored at refrigerated temperature, at room temperature, and at elevated temperatures tend to decrease in ethanol concentration with storage. It appears that the stability of ethanol in blood exposed to temperatures greater than 38°C has not been evaluated. The case presented here involves comparison of breath test results with subsequent analysis of blood drawn at the time of breath testing. However, the blood tubes were in a refrigerator fire followed by refrigerated storage for 5 months prior to analysis by headspace gas chromatography. The subject’s breath was tested twice using an Intoxilyzer 8000. The subject’s blood was tested in duplicate using an Agilent headspace gas chromatograph. The measured breath ethanol concentration was 0.103 g/210 L and 0.092 g/210 L. The measured blood ethanol concentration was 0.0932 g/dL for both samples analyzed. Although the mean blood test result was slightly lower than the mean breath test result, the mean breath test result was within the estimated uncertainty of the mean blood test result. Even under the extreme conditions of the blood kit being in a refrigerator fire, the measured blood ethanol content agreed well with the paired breath ethanol test.     

The unreliability of using a urine ethanol concentration to predict a blood ethanol concentration

Of approximately 5,000 forensic cases with a positive ethanol result, over 1,000 were available in which both blood and urine were present for comparison of ethanol content. Data were examined for calculation of the urine to blood ethanol concentration ratio, with the intent of evaluating the validity of predicting a blood ethanol level given a urine ethanol level. The overall urine to blood ethanol concentration ratio was 1.57:1 with a range of 0.7 to 21.0:1. The extremely wide range of values implies that a large degree of error would be introduced if a mean ratio was used when predicting a blood ethanol level from a urine ethanol level.     

Experimental studies on the mechanism of ethanol formation in corpses

Various in vitro experiments were performed for the purpose of clarifying the mechanism of ethanol production in corpses. Whereas a negligible quantity of ethanol was produced in the blood alone, which was left at room temperature, the quantity of ethanol was slightly increased by addition of glucose to the blood. When saprogens were further added, the quantity was markedly increased. Various materials were added to blood-liver homogenates as specimens, and the mixtures were stored in an incubator at 37 degrees C. As a result of the addition of an antibiotic to the mixture every day, there was hardly any production of ethanol. When alcohol dehydrogenase (ADH) and reduced nicotinamide adenine dinucleotide (NADH) were added, ethanol production was slightly increased. When acetaldehyde was added first, ethanol production was inhibited the next day, but on and after day 2, the quantity of ethanol was more than that in the control material. When pyruvic acid was added first, the results were similar to the above. Pyrazole, cyanamide, and disulfiram completely inhibited the production of ethanol. Ethanol production in corpses is believed to take place through a pathway opposite to that of ethanol metabolism in the living body, under the influence of ADH, ALDH, etc., in saprogens using carbohydrates as substrates.     

Delayed ethanol analysis of breath specimens: long-term field experience with commercial silica gel tubes and breathalyzer collection

Delayed ethanol analysis was performed on breath specimens collected with commercial silica gel tubes using multiple Breathalyzer instruments. Eleven hundred and nine results were obtained from an ethanol testing program over a five-year period. Only 2.5% of the specimens had apparent collection errors. For the valid specimens, the most frequent result was 0.11 g/210 L and the mean result was 0.14 g/210 L. For 642 specimens, delayed results were compared with direct results. Direct results were greater than delayed results for 55%, less than for 27%, and equal to for 18% of the pairs. When fixed tolerance limits of +/- 0.03 were used, 81% of the direct results were confirmed. The confirmation percentage was best in the critical range of direct results, 0.05 to 0.15 g/210 L. The collection tubes showed no substantial variability in retaining ethanol during storage and releasing ethanol for analysis.     

Utilizing the urinary 5-HTOL/5-HIAA ratio to determine ethanol origin in civil aviation accident victims

Specimens from fatal aviation accident victims are submitted to the FAA Civil Aerospace Medical Institute for toxicological analysis. During toxicological evaluations, ethanol analysis is performed on all cases. Care must be taken when interpreting a positive ethanol result due to the potential for postmortem ethanol formation. Several indicators of postmortem ethanol formation exist; however, none are completely reliable. The consumption of ethanol has been shown to alter the concentration of two major serotonin metabolites, 5-hydroxytryptophol (5-HTOL) and 5-hydroxyindole-3-acetic acid (5-HIAA). While the 5-HTOL/5-HIAA ratio is normally very low, previous studies using living subjects have demonstrated that the urinary 5-HTOL/5-HIAA ratio is significantly elevated for 11-19 h after acute ethanol ingestion. Recently, our laboratory developed and validated an analytical method for the simultaneous determination of both 5-HTOL and 5-HIAA in forensic urine samples using a simple liquid/liquid extraction and LC/MS/MS and LC/MS/MS/MS. In this previous work a 15 pmol/nmol serotonin metabolite ratio cutoff was established in postmortem urine, below which it could be conclusively determined that no recent antemortem ethanol consumption had occurred. In the current study this newly validated analytical method was applied to five ethanol-positive aviation fatalities where the origin of the ethanol present could not previously be conclusively determined. In four of the five cases examined the detected ethanol was demonstrated to be present due to postmortem microbial formation, and not consumption, even though some indication of ethanol consumption may have been present.     

A fatal interaction of methocarbamol and ethanol in an accidental poisoning

A case is presented of a fatal drug interaction caused by ingestion of methocarbamol (Robaxin) and ethanol. Methocarbamol is a carbamate derivative used as a muscle relaxant with sedative effects. Therapeutic concentrations of methocarbamol are reported to be 24 to 41 micrograms/mL. Biological fluids were screened for ethanol using the Abbott TDx system and quantitated by gas-liquid chromatography (GLC). Determination of methocarbamol concentrations in biological tissue homogenates and fluids were obtained by colorimetric analysis of diazotized methocarbamol. Blood ethanol concentration was 135 mg/dL (0.135% w/v) and urine ethanol was 249 mg/dL (0.249% w/v). Methocarbamol concentrations were: blood, 257 micrograms/mL; bile, 927 micrograms/L; urine, 255 micrograms/L; gastric, 3.7 g; liver, 459 micrograms/g; and kidney, 83 micrograms/g. The combination of ethanol and carbamates is contraindicated since acute alcohol intoxication combined with carbamate usage can lead to combined central nervous system depression as a result of the interactive sedative-hypnotic properties of the compounds.     

An unusual form of fatal ethanol intoxication

Forensic pathologists are very familiar with deaths due to ethanol intoxication. The overwhelming majority of these deaths are a result of the oral ingestion of ethanol. We report an unusual case of an individual who expired in his secured residence after self administration of a wine enema. Toxicology showed an ethanol concentration of 0.40 g/dL in the blood and 0.41 g/dL in the vitreous fluid. Scene investigation was of paramount importance in determining the unusual method by which the decedent absorbed the alcoholic beverage.     

自动顶空GC/MS测定血液中乙醇含量不确定度评定

目的评定自动顶空—气相色谱—质谱法(GC/MS)测定血液中乙醇含量的不确定度。方法从分析测量过程着手,依据不确定度评定的指导性文件,分析了不确定度来源,量化不确定度分量,计算检测结果的合成标准不确定度和扩展不确定度。结果血液样本两次测定结果平均值为0.738mg/mL的扩展不确定度为0.084mg/mL。结论血液中乙醇含量的不确定度主要来源于样品检测、乙醇标准溶液和标准曲线。