甲氰菊酯法医毒物动力学研究
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摘要
目的:
1.建立家兔甲氰菊酯灌胃染毒死后弥散模型;
2.建立犬甲氰菊酯灌胃染毒模型;
3.改进甲氰菊酯气相色谱检测,建立其GC-ECD、气相色谱/质谱联用检测方法;
4.研究甲氰菊酯在家兔体内的毒物动力学、死后弥散及其在犬体内的分布。
方法:
1.甲氰菊酯在家兔体内的毒物动力学研究取家兔8只,随机分为实验组(6只),分别经口灌胃染毒1/2 LD_(50)甲氰菊酯农药,于15、30、60、120、240和360分钟抽取动脉血1毫升,样品经酸处理后,三氯甲烷萃取,气相色谱和气相色谱.质谱联用检测,根据甲氰菊酯的特征离子峰、保留时间定性与内标法、工作曲线法定量。
2.甲氰菊酯在犬体内的分布研究取犬8只,随机分为实验组(5只),分别经口灌胃染毒8LD_(50)甲氰菊酯农药,2.5小时后将犬处死,迅速解剖动物,取全部心、肝、脾、肺、肾、脑、胃壁、心血、胆汁、胸肌、右下肢肌肉等组织,放置于-20℃冰箱内保存待检,样品经酸处理后,三氯甲烷萃取,气相色谱和气相色谱.质谱联用检测,根据甲氰菊酯的特征离子峰、保留时间定性与内标法、工作曲线法定量,研究犬体内的甲氰菊酯分布情况。
3.甲氰菊酯在家兔体内的死后弥散研究取家兔39只,分为死后弥散组、温度影响组和剂量影响组。死后弥散组:21只,将家兔处死,1小时后,经口灌胃染毒2 LD_(50)甲氰菊酯农药,置于常温环境下,分别于3、6、12、24、48、72和96小时各解剖3只家兔,检测比较家兔脏器中甲氰菊酯的含量;温度影响组:9只,将家兔处死,1小时后经口灌胃染毒2LD_(50)甲氰菊酯农药,每3只置于-20℃、4℃和常温环境下72小时,检测比较家兔脏器中甲氰菊酯的含量;剂量影响组:9只,将家兔处死,1小时后,每3只家兔分别经口灌胃染毒1/2 LD_(50)、2LD_(50)和4 LD_(50)甲氰菊酯农药,置于常温环境下72小时,检测比较家兔脏器中甲氰菊酯的含量。
结果:
1.甲氰菊酯在家兔体内的毒物动力学符合一级动力学、一级吸收、一室开放模型,吸收相和消除相半衰期分别为22分钟和287分钟。
2.犬8LD_(50)剂量甲氰菊酯灌胃染毒后0.95±0.72小时出现大小便失禁,痉挛,角膜反射减弱、呼吸缓慢,心率降低等中毒症状:染毒后2.5h犬体内甲氰菊酯的平均含量从高到低依次为:胃、肝、心血、心、脾、脑、肾、尿、肺、胸肌、右下肢肌肉、玻璃体、胆汁。
3.家兔处死后1小时2LD_(50)甲氰菊酯灌胃染毒3小时后,心血、心、脾、尿和肺中均检出甲氰菊酯,6、12、24、48、72和96小时后各脏器和体液中均检出甲氰菊酯;死后染毒72小时,家兔脏器中甲氰菊酯的含量染毒剂量增大(1/2 LD_(50)、2LD_(50)和4 LD_(50))各脏器中甲氰菊酯检出浓度增加:死后染毒.20℃保存72小时,脑、肝、肾、心、肺、脾中检出甲氰菊酯,胸肌、右下肢肌肉、心血、尿、玻璃体、胆汁中未检出甲氰菊酯;死后染毒4℃保存72小时,仅玻璃体液和胆汁中未检出甲氰菊酯。
4.本研究建立的气相色谱检测生物样本中甲氰菊酯的方法,达到良好的分离,线性范围为2-120μg/g或μg/ml,最低检出浓度是0.013μg/g或μg/ml,回收率均在80%以上。
结论:
1.本研究建立的家兔甲氰菊酯灌胃染毒及死后弥散模型和犬甲氰菊酯灌胃染毒模型可应用于甲氰菊酯意外死亡和中毒案件的法医学研究。
2.本研究建立了甲氰菊酯的GC-ECD检测方法,可应用于甲氰菊酯中毒死亡的法医学检验和法医毒物动力学研究。
3.犬8 LD_(50)染毒后2.5小时犬体内甲氰菊酯的平均含量从高到低依次为:胃、肝、血、心、脾、脑、肾、尿、肺、胸肌、右下肢、玻璃体、胆汁。
4.甲氰菊酯在家兔体内可发生死后弥散,可能与甲氰菊酯本身理化性质、染毒剂量、毒物入体方式、死后给药时间、尸体保存时间和温度、弥散距离、尸体姿势有关。
5.甲氰菊酯中毒死亡法医学鉴定中,除提取胃内容物、肝脏、心血等常见检材外,还应采取肾、脾、脑等组织进行全面的定性和定量分析,并充分考虑药物剂量、死亡时间,尸体所处环境、体内微生物等影响因素,结合临床表现、病理报告等进行综合分析,为甲氰菊酯误服、中毒死亡案件和死后灌毒案件的甄别提供科学、客观的依据。
OBJECTIVE:1.To establish a postmortem diffusion model and a toxicokinetic model of fenpropathrin with rabbits.
2.To establish an intragastric administration model of fenpropathrin in dogs.
3.To develop a GC equipped with an ECD and a GC-MS analysis for fenpropathrin determination.
4.To investigate the toxicokinetics and postmortem diffusion of fenpropathrin in rabbits and to investigate the distribution of fenpropathrin in dogs.
METHODS:1.Study on the toxicokinetics of fenpropathrin in rabbits:Eight rabbits were randomly allocated to one group(n=6 per group ),six rabbits were given fenpropathrin intragastric administration with 0.5LD_(50),1ml arterial blood was drawn out in 15、30、60、120、240 and 360min.After acid treatment these obtained samples were extracted by trichlormethane. Analysis was performed with a GC equipped with an ECD and a GC-MS.The analyses identification was based on retention time in the chromatographic system coupled with the ion fragmentation spectrum in the mass spectrometer.The quantitative analysis was on an internal standard method.
2.Study on the diffusion of fenpropathrin in dogs:Five dogs were given fenpropathrin intragastric administration with 8LD_(50),executed after 2.5 hour,dissected immediately and heart, liver,lien,lung,kidney,brain,gastric wall,heart-blood,bile,breast muscle,muscle of right lower limbs were collected and preserved in -20℃.After acid treatment these obtained samples were extracted by trichlormethane.Analysis was performed with a GC equipped with an ECD and a GC-MS.The analyses identification was based on retention time in the chromatographic system coupled with the ion fragmentation spectrum in the mass spectrometer.The quantitative analysis was on an internal standard method.
3.Study on the postmortem diffusion of fenpropathrin in rabbits:Thirty nine rabbits were randomly allocated to temperature group(n=9 per group,9 rabbits were executed,and given fenpropathrin intragastric administration with 2LD_(50) after 1.5 hour,every 3 rabbits were in -20℃,4℃and normal temperature for 72 hour),dose group(n=9 per group,9 rabbits were executed,and every 3 rabbits were given fenpropathrin intragastric administration with 0.5 LD_(50),2LD_(50) and 4LD_(50) after 1.5 hour,9 rabbits were in normal temperature for 72 hour) and postmortem diffusion time group(n=21 per group,21 rabbits were executed and were given fenpropathrin intragastric administration with 2LD_(50) after 1.5 hour,21 rabbits were in normal temperature,every 3 rabbits were anatomized after 3,6,12,24,48,72 and 96 hour).
RESULTS:1.The toxicokinetics of fenpropathfin in rabbits meets a first order kinetics, first order absorption,one compartment open model.The half time of absorption phase and elimination phase are 22min and 287min.
2.Urinary and fecal incontinence,spasm,decreased corneal reflex,muscular tension decrease and dyspnoea occurred at 0.95±0.72 hour after an intragastric administration with fenpropathrin.Analysis showed that the distribution of fenpropathfin in dogs:the average content of fenpropathrin from high to low:gastric,liver,heart-blood,heart,lien,brain,kidney, urine,lung,breast muscle,muscle of right lower limbs,vitreous and bile.
3.The result of fenpropathrin intragastric postmortem diffusion of rabbits showed that fenpropathrin were detected in heart-blood,heart,lien,urine and lung after the postmortem administration for 3h,and in all organ and body fluid after the postmortem administration for 6, 12,24,48,72 and 96.Dose and preserved temperature related to the postmortem diffusion of fenpropathrin in rabbits.
4.The study has developed a GC equipped with an ECD and a GC-MS analysis for fenpropathrin.The linear range is 2-120μg/g orμg/ml,the minimum detectable concentration is 0.013μg/g orμg/ml,and the average of recovery rate exceeds 80%.
CONCLUSION:1.The toxicokinetics model and postmortem diffusion model developed in this paper can be applied to the forensic identification and study of fenpropathrin poisoning death.
2.The GC-ECD method can be used in the forensic identification and forensic toxicokinetics study of fenpropathrin poisoning death.
3.The order of the fenpropathrin concentration detected in dogs after a 8 LD50 dose of fenpropathrin for 2.5h are:stomach,liver,blood,heart,spleen,brain,kidney,urine,lung,chest muscle,right lower limb muscle,humor vitreous,bile.
4.There is a postmortem diffusion of fenpropathrin in the rabbits,which probably relates to its physico-chemical property,dose,poisoning way,poisoning time after the death,the temperature and time of the corpse to preserve,diffusion distance,corpse posture.
5.Beside common samples such as gastric contents,liver and blood,more samples such as kidney,spleen and brain should be obtained for qualitative and quantitative analysis in the forensic identification of fenpropathrin poisoning death,the dose,death time,environment and micro-organism in vivo should be taken into consideration.
1.建立家兔甲氰菊酯灌胃染毒死后弥散模型;
2.建立犬甲氰菊酯灌胃染毒模型;
3.改进甲氰菊酯气相色谱检测,建立其GC-ECD、气相色谱/质谱联用检测方法;
4.研究甲氰菊酯在家兔体内的毒物动力学、死后弥散及其在犬体内的分布。
方法:
1.甲氰菊酯在家兔体内的毒物动力学研究取家兔8只,随机分为实验组(6只),分别经口灌胃染毒1/2 LD_(50)甲氰菊酯农药,于15、30、60、120、240和360分钟抽取动脉血1毫升,样品经酸处理后,三氯甲烷萃取,气相色谱和气相色谱.质谱联用检测,根据甲氰菊酯的特征离子峰、保留时间定性与内标法、工作曲线法定量。
2.甲氰菊酯在犬体内的分布研究取犬8只,随机分为实验组(5只),分别经口灌胃染毒8LD_(50)甲氰菊酯农药,2.5小时后将犬处死,迅速解剖动物,取全部心、肝、脾、肺、肾、脑、胃壁、心血、胆汁、胸肌、右下肢肌肉等组织,放置于-20℃冰箱内保存待检,样品经酸处理后,三氯甲烷萃取,气相色谱和气相色谱.质谱联用检测,根据甲氰菊酯的特征离子峰、保留时间定性与内标法、工作曲线法定量,研究犬体内的甲氰菊酯分布情况。
3.甲氰菊酯在家兔体内的死后弥散研究取家兔39只,分为死后弥散组、温度影响组和剂量影响组。死后弥散组:21只,将家兔处死,1小时后,经口灌胃染毒2 LD_(50)甲氰菊酯农药,置于常温环境下,分别于3、6、12、24、48、72和96小时各解剖3只家兔,检测比较家兔脏器中甲氰菊酯的含量;温度影响组:9只,将家兔处死,1小时后经口灌胃染毒2LD_(50)甲氰菊酯农药,每3只置于-20℃、4℃和常温环境下72小时,检测比较家兔脏器中甲氰菊酯的含量;剂量影响组:9只,将家兔处死,1小时后,每3只家兔分别经口灌胃染毒1/2 LD_(50)、2LD_(50)和4 LD_(50)甲氰菊酯农药,置于常温环境下72小时,检测比较家兔脏器中甲氰菊酯的含量。
结果:
1.甲氰菊酯在家兔体内的毒物动力学符合一级动力学、一级吸收、一室开放模型,吸收相和消除相半衰期分别为22分钟和287分钟。
2.犬8LD_(50)剂量甲氰菊酯灌胃染毒后0.95±0.72小时出现大小便失禁,痉挛,角膜反射减弱、呼吸缓慢,心率降低等中毒症状:染毒后2.5h犬体内甲氰菊酯的平均含量从高到低依次为:胃、肝、心血、心、脾、脑、肾、尿、肺、胸肌、右下肢肌肉、玻璃体、胆汁。
3.家兔处死后1小时2LD_(50)甲氰菊酯灌胃染毒3小时后,心血、心、脾、尿和肺中均检出甲氰菊酯,6、12、24、48、72和96小时后各脏器和体液中均检出甲氰菊酯;死后染毒72小时,家兔脏器中甲氰菊酯的含量染毒剂量增大(1/2 LD_(50)、2LD_(50)和4 LD_(50))各脏器中甲氰菊酯检出浓度增加:死后染毒.20℃保存72小时,脑、肝、肾、心、肺、脾中检出甲氰菊酯,胸肌、右下肢肌肉、心血、尿、玻璃体、胆汁中未检出甲氰菊酯;死后染毒4℃保存72小时,仅玻璃体液和胆汁中未检出甲氰菊酯。
4.本研究建立的气相色谱检测生物样本中甲氰菊酯的方法,达到良好的分离,线性范围为2-120μg/g或μg/ml,最低检出浓度是0.013μg/g或μg/ml,回收率均在80%以上。
结论:
1.本研究建立的家兔甲氰菊酯灌胃染毒及死后弥散模型和犬甲氰菊酯灌胃染毒模型可应用于甲氰菊酯意外死亡和中毒案件的法医学研究。
2.本研究建立了甲氰菊酯的GC-ECD检测方法,可应用于甲氰菊酯中毒死亡的法医学检验和法医毒物动力学研究。
3.犬8 LD_(50)染毒后2.5小时犬体内甲氰菊酯的平均含量从高到低依次为:胃、肝、血、心、脾、脑、肾、尿、肺、胸肌、右下肢、玻璃体、胆汁。
4.甲氰菊酯在家兔体内可发生死后弥散,可能与甲氰菊酯本身理化性质、染毒剂量、毒物入体方式、死后给药时间、尸体保存时间和温度、弥散距离、尸体姿势有关。
5.甲氰菊酯中毒死亡法医学鉴定中,除提取胃内容物、肝脏、心血等常见检材外,还应采取肾、脾、脑等组织进行全面的定性和定量分析,并充分考虑药物剂量、死亡时间,尸体所处环境、体内微生物等影响因素,结合临床表现、病理报告等进行综合分析,为甲氰菊酯误服、中毒死亡案件和死后灌毒案件的甄别提供科学、客观的依据。
OBJECTIVE:1.To establish a postmortem diffusion model and a toxicokinetic model of fenpropathrin with rabbits.
2.To establish an intragastric administration model of fenpropathrin in dogs.
3.To develop a GC equipped with an ECD and a GC-MS analysis for fenpropathrin determination.
4.To investigate the toxicokinetics and postmortem diffusion of fenpropathrin in rabbits and to investigate the distribution of fenpropathrin in dogs.
METHODS:1.Study on the toxicokinetics of fenpropathrin in rabbits:Eight rabbits were randomly allocated to one group(n=6 per group ),six rabbits were given fenpropathrin intragastric administration with 0.5LD_(50),1ml arterial blood was drawn out in 15、30、60、120、240 and 360min.After acid treatment these obtained samples were extracted by trichlormethane. Analysis was performed with a GC equipped with an ECD and a GC-MS.The analyses identification was based on retention time in the chromatographic system coupled with the ion fragmentation spectrum in the mass spectrometer.The quantitative analysis was on an internal standard method.
2.Study on the diffusion of fenpropathrin in dogs:Five dogs were given fenpropathrin intragastric administration with 8LD_(50),executed after 2.5 hour,dissected immediately and heart, liver,lien,lung,kidney,brain,gastric wall,heart-blood,bile,breast muscle,muscle of right lower limbs were collected and preserved in -20℃.After acid treatment these obtained samples were extracted by trichlormethane.Analysis was performed with a GC equipped with an ECD and a GC-MS.The analyses identification was based on retention time in the chromatographic system coupled with the ion fragmentation spectrum in the mass spectrometer.The quantitative analysis was on an internal standard method.
3.Study on the postmortem diffusion of fenpropathrin in rabbits:Thirty nine rabbits were randomly allocated to temperature group(n=9 per group,9 rabbits were executed,and given fenpropathrin intragastric administration with 2LD_(50) after 1.5 hour,every 3 rabbits were in -20℃,4℃and normal temperature for 72 hour),dose group(n=9 per group,9 rabbits were executed,and every 3 rabbits were given fenpropathrin intragastric administration with 0.5 LD_(50),2LD_(50) and 4LD_(50) after 1.5 hour,9 rabbits were in normal temperature for 72 hour) and postmortem diffusion time group(n=21 per group,21 rabbits were executed and were given fenpropathrin intragastric administration with 2LD_(50) after 1.5 hour,21 rabbits were in normal temperature,every 3 rabbits were anatomized after 3,6,12,24,48,72 and 96 hour).
RESULTS:1.The toxicokinetics of fenpropathfin in rabbits meets a first order kinetics, first order absorption,one compartment open model.The half time of absorption phase and elimination phase are 22min and 287min.
2.Urinary and fecal incontinence,spasm,decreased corneal reflex,muscular tension decrease and dyspnoea occurred at 0.95±0.72 hour after an intragastric administration with fenpropathrin.Analysis showed that the distribution of fenpropathfin in dogs:the average content of fenpropathrin from high to low:gastric,liver,heart-blood,heart,lien,brain,kidney, urine,lung,breast muscle,muscle of right lower limbs,vitreous and bile.
3.The result of fenpropathrin intragastric postmortem diffusion of rabbits showed that fenpropathrin were detected in heart-blood,heart,lien,urine and lung after the postmortem administration for 3h,and in all organ and body fluid after the postmortem administration for 6, 12,24,48,72 and 96.Dose and preserved temperature related to the postmortem diffusion of fenpropathrin in rabbits.
4.The study has developed a GC equipped with an ECD and a GC-MS analysis for fenpropathrin.The linear range is 2-120μg/g orμg/ml,the minimum detectable concentration is 0.013μg/g orμg/ml,and the average of recovery rate exceeds 80%.
CONCLUSION:1.The toxicokinetics model and postmortem diffusion model developed in this paper can be applied to the forensic identification and study of fenpropathrin poisoning death.
2.The GC-ECD method can be used in the forensic identification and forensic toxicokinetics study of fenpropathrin poisoning death.
3.The order of the fenpropathrin concentration detected in dogs after a 8 LD50 dose of fenpropathrin for 2.5h are:stomach,liver,blood,heart,spleen,brain,kidney,urine,lung,chest muscle,right lower limb muscle,humor vitreous,bile.
4.There is a postmortem diffusion of fenpropathrin in the rabbits,which probably relates to its physico-chemical property,dose,poisoning way,poisoning time after the death,the temperature and time of the corpse to preserve,diffusion distance,corpse posture.
5.Beside common samples such as gastric contents,liver and blood,more samples such as kidney,spleen and brain should be obtained for qualitative and quantitative analysis in the forensic identification of fenpropathrin poisoning death,the dose,death time,environment and micro-organism in vivo should be taken into consideration.
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