苯巴比妥的法医毒物动力学研究
发布时间：2018-09-08 11:41
【摘要】： 目的 1.建立生物检材中苯巴比妥的气相色谱和气相色谱/质谱检测方法; 2.建立苯巴比妥的毒物动力学、动态分布、死后分布、死后再分布动物模型; 3.研究苯巴比妥在大鼠体内的毒物动力学、动态分布、死后分布、死后再分布规律,为苯巴比妥中毒(死)案件的法医学鉴定提供实验依据。 方法 1.样品的处理和气相色谱、气相色谱/质谱检测:血、组织样品中加入内标物丙烯巴比妥,经盐酸酸化后(pH=1～2),乙醚萃取,气相色谱/质谱联用法定性,气相色谱内标法定量检测苯巴比妥。 2.苯巴比妥在急性中毒大鼠体内的毒物动力学及动态分布的研究:雄性wistar大鼠54只,经灌胃匀速注入1/2LD50苯巴比妥(330 mg·kg~(-1)),给药后观察大鼠的中毒症状,分别于给药后1、5、10、15、20、30、48、72、96h各处死6只,迅速解剖动物,取心血、心、肝、脾、肺、肾、脑等冷冻保存,酸性乙醚提取,气相色谱质谱法定性、气相色谱定量检测其中苯巴比妥。 3.苯巴比妥在大鼠体内的死后分布研究:雄性wistar大鼠12只,随机分LD50组和2LD50组,经灌胃匀速注入LD50(0.66 mg·kg~(-1))和2LD50(1.32 mg·kg~(-1))苯巴比妥。观察给药到死亡时的生命体征的变化以及中毒症状,待呼吸和心跳全部消失时,迅速解剖动物,取心血、心、肝、脾、肺、肾、脑、胃等冷冻保存,酸性乙醚提取,气相色谱质谱法定性、气相色谱定量检测苯巴比妥。 4.苯巴比妥在大鼠体内的死后再分布研究:雄性wistar大鼠54只,以1/2LD50(330 mg·kg~(-1))灌胃,10小时后处死,室温(20?C)条件下,于死后0、2、4、8、12、24、48、72、96小时取心血、心、肝、脾、肺、肾、大脑、肌肉,酸性乙醚提取,气相色谱质谱法定性、气相色谱定量检测其中苯巴比妥。 5.统计学方法:采用SPSS11.5统计软件处理数据,结果以均数±标准差( )表示,t检验。 结果 1.气相色谱和气相色谱/质谱检测:气相色谱/质谱分析苯巴比妥的特征离子m/z为204、232。心血中苯巴比妥气相色谱检测的回归方程、线性检测范围、相关系数、回收率、最低检出浓度分别为Y = 43.476X + 0.8168(μg·mL~(-1))、0.5 -280μg·mL~(-1)、0.9955、97.50±2.0%、0.5μg·mL~(-1);肝组织中苯巴比妥气相色谱检测的回归方程、线性检测范围、相关系数、回收率、最低检出浓度分别为Y = 46.867X + 0.0884 (μg·g~(-1))、0.5 -280μg·g~(-1)、0.994、98.5±3.5%、0.5μg·g~(-1)。 2.毒物动力学和动态分布:1/2LD50剂量苯巴比妥在大鼠体内的毒物动力学符合口服给药一级动力学一室开放模型过程,各脏器中苯巴比妥的毒物动力学符合血管外给药一级动力学一室开放模型。心血中达峰时间和消除半衰期除为7.39h和26.02h;各脏器中达峰时间和消除半衰期为2.41h~4.73h和8.59h~20.91h;经口染毒后不同时间大鼠体内苯巴比妥含量顺序为肝、心血、心和肺、其他组织,各脏器内苯巴比妥含量在5h左右达高峰,之后呈下降趋势。 3.死后分布:大鼠在苯巴比妥灌胃后活动明显减少,出现嗜睡症状,LD50灌胃组大鼠7.24±3.17h死亡,2LD50灌胃组大鼠1.33±0.25h死亡。各脏器组织苯巴比妥含量由高到低分别为:①LD50组:胃(1261.34±591.83)心(332.12±70.87)、脑(289.81±71.43)、肝(284.68±79.44)、肺(284.41±67.47)、脾(272.56±31.53)、心血(269.52±44.62)肾(167.76±43.72);②2LD50组:胃(1649.21±1212.82)肝(357.83±138.67)、脑(323.19±159.92)、心(310.95±130.73)、心血(309.93±135.60)、肺(308.28±134.48)、脾(299.77±104.58)肾(233.24±91.92)。 4.死后再分布:大鼠在处死后96小时内,在室温(20?C)条件下,死后2~4小时的心血、心、肺、肾、脑、肌的苯巴比妥含量与死亡当时含量相比明显升高(p0.05),而后逐渐降低;死后2小时肝、脾的苯巴比妥含量与死亡当时含量相比明显升高(p0.05),则出现逐渐升高的趋势(p0.05)。 结论 1.本文建立的生物检材中苯巴比妥气相色谱/质谱检测方法选择性好,定性准确,气相色谱检测简便,快速,灵敏,定量结果准确,可用于苯巴比妥中毒的临床快速检验诊断和苯巴比妥中毒(死)案件的法医学鉴定。 2.本文建立了苯巴比妥的毒物动力学、动态分布、死后分布、死后再分布研究动物模型,可应用于苯巴比妥的法医毒物动力学研究。 3.苯巴比妥在大鼠血和各脏器中毒物动力学均符合血管外给药一级动力学一室开放模型。急性染毒大鼠体内苯巴比妥含量顺序为,肝、心血、心和肺、其他组织,各脏器内苯巴比妥含量在5h左右达高峰,之后呈下降趋势。 4.苯巴比妥死后分布不均匀,含血丰富和脂质含量高的器官如肝、肺、心、脑较其它组织和血液含量高。法医鉴定苯巴比妥中毒(死)案件时,应全面正确的采取检材进行毒物分析。 5.苯巴比妥在染毒大鼠体内可发生死后再分布,死后心血、心、肝、脾、肺、肾、脑和肌中苯巴比妥浓度均出现升高。
[Abstract]:objective
1. establish a gas chromatography and gas chromatography-mass spectrometry (GC / MS) method for the detection of phenobarbital in biological samples.
2. establishment of phenobarbital toxicology, dynamic distribution, postmortem distribution, postmortem redistribution animal model.
3. To study the toxicokinetics, dynamic distribution, postmortem distribution and postmortem redistribution of phenobarbital in rats, and to provide experimental basis for forensic identification of phenobarbital poisoning cases.
Method
1. Sample treatment and gas chromatography, gas chromatography/mass spectrometry detection: blood, tissue samples with the addition of internal standard propylene barbiturate, acidified by hydrochloric acid (pH=1-2), ether extraction, gas chromatography/mass spectrometry qualitative, gas chromatography internal standard quantitative detection of phenobarbital.
2. Study on toxicokinetics and dynamic distribution of phenobarbital in rats with acute poisoning: 54 male Wistar rats were injected with 1/2 LD50 phenobarbital (330 mg (-1)) by gastric perfusion at a uniform rate. The toxic symptoms of rats were observed after administration. Six rats were executed at 1,5,10,15,20,30,48,72,96 h after administration, respectively. Spleen, lung, kidney, brain and other cryopreservation, acid ether extraction, gas chromatography-mass spectrometry, gas chromatography-quantitative detection of phenobarbital.
3. Study on postmortem distribution of phenobarbital in rats: 12 male Wistar rats were randomly divided into LD50 group and 2LD50 group. LD50 (0.66 mg (-1)) and 2LD50 (1.32 mg (-1)) phenobarbital were injected into the stomach at a uniform rate. The changes of vital signs and poisoning symptoms at the time of death were observed and the respiratory and heartbeat symptoms disappeared. Animals, blood, heart, liver, spleen, lung, kidney, brain, stomach and other cryopreservation, acid ether extraction, gas chromatography-mass spectrometry, gas chromatography quantitative detection of phenobarbital.
4. Study on postmortem redistribution of phenobarbital in rats: 54 male Wistar rats were orally administered with 1/2 LD50 (330 mg (-1)) for 10 hours and then executed at room temperature (20?C). Cardiac blood, heart, liver, spleen, lung, kidney, brain, muscle, acidic ether were collected at 0, 2, 4, 8, 12, 24, 48, 72, 96 hours after death. Phenobarbital was detected.
5. Statistical methods: SPSS11.5 statistical software was used to process the data. The results were expressed as mean [standard deviation], t test.
Result
1. Gas Chromatography and Gas Chromatography/Mass Spectrometry: Characteristic ion m/z of phenobarbital was 204,232. Regression equation, linear detection range, correlation coefficient, recovery, minimum detection concentration Y=43.476X+0.8168 (0.5-280ug.mL~(-1), 0.9955, 97. The regression equation, linear detection range, correlation coefficient and recovery rate of phenobarbital gas chromatography in liver tissue were Y=46.867X+0.0884(ug.g~(-1)), 0.5-280ug.g~(-1), 0.994,98.5 (-3.5%) and 0.5ug.g~(-1)), respectively.
2. Toxicokinetics and Dynamic Distribution: The toxicokinetics of 1/2 LD50 dose of phenobarbital in rats conformed to the one-compartment open model of oral administration. The toxicokinetics of phenobarbital in organs conformed to the one-compartment open model of extravascular administration. The peak time and elimination half-life were 2.41h~4.73h and 8.59h~20.91h in each organ, and the phenobarbital content in liver, heart blood, heart and lung, other tissues and organs reached the peak at about 5h, then decreased.
3. Distribution of postmortem: The activity of rats decreased significantly after intragastric administration of phenobarbital, and the symptoms of somnolence were observed. The rats in LD50 group died at 7.24 [3.17 hours] and the rats in 2LD50 group died at 1.33 [0.25 hours] respectively. The contents of phenobarbital in various organs and tissues were: (1) LD50 group: gastric (1261.34 [591.83] heart (332.12 [70.87], brain (289.81 [71.43]), liver (284.68 [79.44]] respectively. (2) 2LD50 group: stomach (1649.21 [1212 12.82) liver (1649.21 [(1649.21 [1212.82), (357.83 [138.67], brain (323.19 [159.92], heart (310.95 [130.73]), heart blood (309.93 [(310.95 [130.73.73], heart (309.93 [309.93 [135.60], heart (308.52 [308.28 [134.28.48].48), spleen (299.77.77 [77.77 [58.58.58.58] kidn (299.58.58.58)] kidney (299.77 3.24 [91.92].
4. Post-death redistribution: Within 96 hours after death, at room temperature (20?C), the phenobarbital content in heart blood, heart, lung, kidney, brain and muscle of rats 2-4 hours after death was significantly higher than that at the time of death (p0.05), and then gradually decreased; 2 hours after death, the phenobarbital content in liver and spleen was significantly higher than that at the time of death (p0.05). The trend is increasing gradually (P0.05).
conclusion
1. The method of gas chromatography/mass spectrometry for phenobarbital in biological samples established in this paper is selective, accurate, simple, rapid, sensitive and accurate. It can be used for rapid clinical diagnosis of phenobarbital poisoning and forensic identification of cases of phenobarbital poisoning (death).
2. In this paper, the toxicokinetics, dynamic distribution, postmortem distribution and postmortem redistribution of phenobarbital were established, which can be used in forensic toxicokinetics of phenobarbital.
3. The toxicokinetics of phenobarbital in rat blood and organs conformed to the one-compartment open model of the first-order kinetics of extravascular administration.
4. The distribution of phenobarbital after death is uneven. The organs with rich blood and high lipid content such as liver, lung, heart and brain are higher than other tissues and blood.
5. Phenobarbital can be redistributed after death in poisoned rats. The concentration of phenobarbital in heart blood, heart, liver, spleen, lung, kidney, brain and muscle increased after death.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919
本文编号：2230412
[Abstract]:objective
1. establish a gas chromatography and gas chromatography-mass spectrometry (GC / MS) method for the detection of phenobarbital in biological samples.
2. establishment of phenobarbital toxicology, dynamic distribution, postmortem distribution, postmortem redistribution animal model.
3. To study the toxicokinetics, dynamic distribution, postmortem distribution and postmortem redistribution of phenobarbital in rats, and to provide experimental basis for forensic identification of phenobarbital poisoning cases.
Method
1. Sample treatment and gas chromatography, gas chromatography/mass spectrometry detection: blood, tissue samples with the addition of internal standard propylene barbiturate, acidified by hydrochloric acid (pH=1-2), ether extraction, gas chromatography/mass spectrometry qualitative, gas chromatography internal standard quantitative detection of phenobarbital.
2. Study on toxicokinetics and dynamic distribution of phenobarbital in rats with acute poisoning: 54 male Wistar rats were injected with 1/2 LD50 phenobarbital (330 mg (-1)) by gastric perfusion at a uniform rate. The toxic symptoms of rats were observed after administration. Six rats were executed at 1,5,10,15,20,30,48,72,96 h after administration, respectively. Spleen, lung, kidney, brain and other cryopreservation, acid ether extraction, gas chromatography-mass spectrometry, gas chromatography-quantitative detection of phenobarbital.
3. Study on postmortem distribution of phenobarbital in rats: 12 male Wistar rats were randomly divided into LD50 group and 2LD50 group. LD50 (0.66 mg (-1)) and 2LD50 (1.32 mg (-1)) phenobarbital were injected into the stomach at a uniform rate. The changes of vital signs and poisoning symptoms at the time of death were observed and the respiratory and heartbeat symptoms disappeared. Animals, blood, heart, liver, spleen, lung, kidney, brain, stomach and other cryopreservation, acid ether extraction, gas chromatography-mass spectrometry, gas chromatography quantitative detection of phenobarbital.
4. Study on postmortem redistribution of phenobarbital in rats: 54 male Wistar rats were orally administered with 1/2 LD50 (330 mg (-1)) for 10 hours and then executed at room temperature (20?C). Cardiac blood, heart, liver, spleen, lung, kidney, brain, muscle, acidic ether were collected at 0, 2, 4, 8, 12, 24, 48, 72, 96 hours after death. Phenobarbital was detected.
5. Statistical methods: SPSS11.5 statistical software was used to process the data. The results were expressed as mean [standard deviation], t test.
Result
1. Gas Chromatography and Gas Chromatography/Mass Spectrometry: Characteristic ion m/z of phenobarbital was 204,232. Regression equation, linear detection range, correlation coefficient, recovery, minimum detection concentration Y=43.476X+0.8168 (0.5-280ug.mL~(-1), 0.9955, 97. The regression equation, linear detection range, correlation coefficient and recovery rate of phenobarbital gas chromatography in liver tissue were Y=46.867X+0.0884(ug.g~(-1)), 0.5-280ug.g~(-1), 0.994,98.5 (-3.5%) and 0.5ug.g~(-1)), respectively.
2. Toxicokinetics and Dynamic Distribution: The toxicokinetics of 1/2 LD50 dose of phenobarbital in rats conformed to the one-compartment open model of oral administration. The toxicokinetics of phenobarbital in organs conformed to the one-compartment open model of extravascular administration. The peak time and elimination half-life were 2.41h~4.73h and 8.59h~20.91h in each organ, and the phenobarbital content in liver, heart blood, heart and lung, other tissues and organs reached the peak at about 5h, then decreased.
3. Distribution of postmortem: The activity of rats decreased significantly after intragastric administration of phenobarbital, and the symptoms of somnolence were observed. The rats in LD50 group died at 7.24 [3.17 hours] and the rats in 2LD50 group died at 1.33 [0.25 hours] respectively. The contents of phenobarbital in various organs and tissues were: (1) LD50 group: gastric (1261.34 [591.83] heart (332.12 [70.87], brain (289.81 [71.43]), liver (284.68 [79.44]] respectively. (2) 2LD50 group: stomach (1649.21 [1212 12.82) liver (1649.21 [(1649.21 [1212.82), (357.83 [138.67], brain (323.19 [159.92], heart (310.95 [130.73]), heart blood (309.93 [(310.95 [130.73.73], heart (309.93 [309.93 [135.60], heart (308.52 [308.28 [134.28.48].48), spleen (299.77.77 [77.77 [58.58.58.58] kidn (299.58.58.58)] kidney (299.77 3.24 [91.92].
4. Post-death redistribution: Within 96 hours after death, at room temperature (20?C), the phenobarbital content in heart blood, heart, lung, kidney, brain and muscle of rats 2-4 hours after death was significantly higher than that at the time of death (p0.05), and then gradually decreased; 2 hours after death, the phenobarbital content in liver and spleen was significantly higher than that at the time of death (p0.05). The trend is increasing gradually (P0.05).
conclusion
1. The method of gas chromatography/mass spectrometry for phenobarbital in biological samples established in this paper is selective, accurate, simple, rapid, sensitive and accurate. It can be used for rapid clinical diagnosis of phenobarbital poisoning and forensic identification of cases of phenobarbital poisoning (death).
2. In this paper, the toxicokinetics, dynamic distribution, postmortem distribution and postmortem redistribution of phenobarbital were established, which can be used in forensic toxicokinetics of phenobarbital.
3. The toxicokinetics of phenobarbital in rat blood and organs conformed to the one-compartment open model of the first-order kinetics of extravascular administration.
4. The distribution of phenobarbital after death is uneven. The organs with rich blood and high lipid content such as liver, lung, heart and brain are higher than other tissues and blood.
5. Phenobarbital can be redistributed after death in poisoned rats. The concentration of phenobarbital in heart blood, heart, liver, spleen, lung, kidney, brain and muscle increased after death.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919
【引证文献】
相关期刊论文 前1条
1 陈跃;朱军;于忠山;喻洪江;刘耀;;毒物代谢动力学研究在法庭科学中的应用[J];中国法医学杂志;2012年06期
相关硕士学位论文 前2条
1 韩亮;苯巴比妥的法医毒物动力学研究（二）[D];山西医科大学;2011年
2 张晓飞;利眠宁的法医毒物动力学研究[D];山西医科大学;2012年
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