本文選題：乙基葡萄糖醛酸苷 + 代謝動力學　； 參考：《復旦大學》2010年碩士論文

【摘要】：我國酒文化源遠流長,因飲酒引發(fā)的事故和案件已經成為嚴重的社會問題。涉及酒精的技術鑒定,如酒后駕駛或醉酒駕駛的認定、酒后肇事逃逸的認定、尸體中酒精來源的判定等,是司法鑒定工作中經常面臨的問題。 乙醇進入人體后,攝入量的95%-98%經肝代謝,先被氧化為乙醛,再進一步被氧化為乙酸；其余的部分(2%-5%)主要以原體形態(tài)由尿液、呼氣和汗液中排出體外；僅有微量乙醒(0.1%)與尿嘧啶核苷-5’-二磷-葡萄糖醛酸結合,生成乙基葡萄糖醛酸苷(Ethyl Glucuronide,Et(?)G)從樣中排出體外；同時有微量乙醇在肝臟中代謝成乙基硫酸酯(Ethyl Sulphate, EtS),由尿液中排出體外。由于乙基葡萄糖醛酸苷和乙基硫酸酯在體內可停留時間比其原體乙醇長,近年來越來越為人們所重視,被認為是乙醇的標志性化合物。在酒后駕車事故,特別是肇事逃逸案件中,當血液中測不出乙醇時,只要檢出一定量的乙基葡萄糖醛酸苷和乙基硫酸酯,也可認為是酒后駕車或生前攝入乙醇。有關乙基葡萄糖醛酸苷的檢測及體內代謝動力學國外已有一些報道,國內尚未有相關研究報道。 本學位論文以乙基葡萄糖醛酸苷為主要研究對象,致力于建立血液和尿液中乙基葡萄糖醛酸苷的定性、定量分析方法,并對方法的檢出限、回收率、線性范圍等作出評價；利用健康自愿受試者控制劑量飲酒后,定時測定其血液和尿液中乙基葡萄糖醛酸苷濃度,考察中國人體內乙基葡萄糖醛酸苷的代謝動力學；確定中國人酒后乙基葡萄糖醛酸苷的檢出時限；初步實現(xiàn)了利用血液中EtG和乙醇濃度關系,推定飲酒時間的數(shù)據(jù)建模。全部內容按照工作展開的時間順序分為六章。 第一章介紹了國內外EtG分析研究進展和應用,闡述了本研究的研究背景、意義和主要工作。 第二章建立和評價了氣相色譜-串聯(lián)質譜聯(lián)用儀(GC-MS/MS)測定血液中EtG的方法。該方法血液中EtG的檢出限(LOD)為0.05μg/mL,線性范圍為0.1μg/mL-10μg/mL (R2=0.9999),準確度為90%-110%,日內精密度和日間精密度均小于15%,并成功應用于實際案例的檢測。 第三章建立和評價了液相色譜-線性離子阱-軌道離子阱(LC-LTQ-Orbitrap)測定血液、尿液中EtG的方法。該方法血液、尿液中EtG的檢測限均為0.02μ/mL,線性范圍為0.05-5.00μg/mL (R20.998),兩基質中EtG檢測方法的日內準確度與日間準確度均為85%-110%,日內精密度和日間精密度均小于10%。 第四章建立和評價了液相色譜-串聯(lián)質譜聯(lián)用儀(LC-MS/MS)測定血液、尿液中EtG的方法。該方法血液和尿液中EtG的檢測限均為0.05μg/mL,線性范圍為0.1μg/mL-5μg/mL (R20.999),兩基質中EtG檢測方法的日內準確度均為95%-105%,日內精密度與日間精密度均小于15%。 第五章開展了劑量控制的健康志愿者飲酒實驗,采集血液與尿液樣本；利用建立的LC-MS/MS方法檢測樣本,考察EtG的代謝動力學,探討了血液和尿液中EtG,血液和尿液中的乙醇的相互關系。通過統(tǒng)計軟件Stata 7.0計算,EtG在血液濃度峰值為4-6h,檢測時限為8-21h,比血液中乙醇檢測時限長8-10h。乙醇在EtG在尿液濃度峰值為4-6h,檢測時限為20-85h,比尿液中乙醇檢測時限長12-70h,并以此初步建立了由EtG和乙醇濃度推算飲酒時間的數(shù)據(jù)模型。 第六章建立了血液、尿液中EtS的LC-LTQ-Orbitrap分析方法。線性范圍0.1μg/mL-5μg/mL (R20.995)。 本研究在國內首次建立了GC-MS/MS、LC-LTQ-Orbitrap和LC-MS/MS等多個EtG檢測方法并進行了方法學驗證,其中LC-LTQ-Orbitrap方法為國際上首次報道；首次考察中國人酒后乙基葡萄糖醛酸苷的代謝動力學,探討了血液和尿液中EtG與乙醇的相互關系,提出了EtG在血液、尿液中的檢出時限。研究成果為酒后駕駛逃逸者的認定、尸體檢材中乙醇來源的判斷提供了科學依據(jù)。
[Abstract]:The wine culture in China has a long history, and the accidents and cases caused by drinking have become serious social problems. The technical identification of alcohol, such as the identification of drunk driving or drunken driving, the identification of the escape and escape after alcohol, the determination of the source of alcohol in the corpse, is a common problem in the judicial appraisal work.
After the ethanol enters the body, the intake of 95%-98% is oxidized to acetaldehyde first and then oxidized to acetic acid; the rest (2%-5%) is mainly discharged from urine, exhaled and perspiration in the form of mycoplasma, and only a trace of awoke (0.1%) is combined with uridine -5 'two phosphorus glucuronic acid to produce ethyl glucuronic acid. Glycosides (Ethyl Glucuronide, Et (?) G) are excreted from the sample in vitro; meanwhile, there are trace amounts of ethanol in the liver that metabolize Cheng Yiji sulfate (Ethyl Sulphate, EtS) and excreted in the urine. As ethyl glucuronide and ethyl sulfate are longer in vivo than their ethanol, more and more attention has been paid to them in recent years. It is a symbol of ethanol. In the case of drunk driving, especially in the accident and escaping case, when a certain amount of ethyl glucuronide and ethyl sulfate are detected in the blood, a certain amount of ethyl glucuronide and ethyl sulfate can be considered to be drunk driving or intake of ethanol in life. There have been some reports outside of China, and no relevant research reports have been reported in China.
This thesis focuses on the determination of ethyl glucuronide in blood and urine, quantitative analysis, detection limit, recovery, linear range, etc. The metabolic kinetics of ethyl glucuronide in Chinese human body was investigated by ethyl glucuronide concentration, and the detection time limit of ethyl glucuronide in Chinese people was determined. The data modeling of using the relationship between the concentration of EtG and ethanol in the blood and determining the time of alcohol consumption was preliminarily realized. The whole content was divided according to the time sequence of work. It is the six chapter.
The first chapter introduces the progress and application of EtG analysis at home and abroad, and expounds the background, significance and main work of this research.
In the second chapter, the method for the determination of EtG in blood by gas chromatography tandem mass spectrometry (GC-MS/MS) is established and evaluated. The detection limit (LOD) of EtG in the blood is 0.05 mu g/mL, the linear range is 0.1 mu g/mL-10 mu g/mL (R2=0.9999), the accuracy is 90%-110%, the intraday precision and the day precision are less than 15%, and are successfully applied to the actual cases. Testing.
In the third chapter, the method for the determination of EtG in blood and urine by liquid chromatography linear ion trap orbital ion trap (LC-LTQ-Orbitrap) is established and evaluated. The detection limit of EtG in blood and urine is 0.02 /mL, and the linear range is 0.05-5.00 mu g/mL (R20.998). The daily accuracy and day accuracy of the two matrix EtG test methods are all 85%-110%, The intraday precision and the day precision are less than 10%.
In the fourth chapter, the method for the determination of EtG in blood and urine by liquid chromatography tandem mass spectrometry (LC-MS/MS) is established and evaluated. The detection limit of EtG in blood and urine is 0.05 g/mL, and the linear range is 0.1 mu g/mL-5 g/mL (R20.999). The intra day accuracy of EtG detection method in the two matrix is all 95%-105%, intraday precision and day precision The degree is less than 15%.
In the fifth chapter, a dose controlled healthy volunteers drinking experiment was carried out to collect blood and urine samples. The metabolic kinetics of EtG was examined by the established LC-MS/MS method. The relationship between the blood and urine in EtG, blood and urine was investigated. The peak of the blood concentration of EtG was calculated by the statistical software Stata 7. The peak of the blood concentration was 4-6 H, the detection time limit is 8-21h, the peak of 8-10h. ethanol at the urine concentration of EtG at EtG is 4-6h, the time limit is 20-85h, and the time limit of the urine ethanol detection is 12-70h, and the data model of calculating the drinking time by the concentration of EtG and ethanol is preliminarily established.
In the sixth chapter, LC-LTQ-Orbitrap analysis of EtS in blood and urine was established. The linear range was 0.1 g/mL-5 g/mL-5 (R20.995).
In this study, multiple EtG detection methods, such as GC-MS/MS, LC-LTQ-Orbitrap and LC-MS/MS, were established and verified for the first time in China. The LC-LTQ-Orbitrap method was first reported in the world. The metabolic kinetics of Chinese alcohol ethyl glucuronide was first investigated, and the relationship between EtG and ethanol in blood and urine was discussed. The time limit for the detection of EtG in the blood and urine was put forward. The research results provided a scientific basis for the identification of the drunk driving runaway and the determination of the sources of ethanol in the corpse inspection.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：D919.2

【參考文獻】

相關期刊論文 前1條

1 沈保華;嚴慧;趙暉;卓先義;;氣相色譜-串聯(lián)質譜法測定血液中乙基葡萄糖醛酸苷[J];法醫(yī)學雜志;2009年06期

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本文編號：1985810