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Objective 1. Establishment of chlorpromazine in biological samples by gas chromatography and gas chromatography / mass spectrometry detection methods; 2. Postmortem distribution of chlorpromazine, Postmortem redistribution after the death of dispersion, the decomposition kinetics animal model; 3. Of chlorpromazine in animals after death distribution Postmortem redistribution, after the death of diffusion and decomposition kinetics of distribution law to provide experimental evidence for chlorpromazine the poisoning (dead) cases forensic identification. The Method 1 death Distribution of 1.1 chlorpromazine in rats in vivo distribution studies after death: six male SD rats, weight 200g ± 10g by intragastric uniform injection of 6LD 50 (1350 mg / kg) of chlorpromazine. Observed changes in vital signs when death and symptoms of poisoning administration to be breathing and heartbeat disappeared quickly anatomy animal, take the cryopreservation of the brain, heart, liver, spleen, lung, kidney, heart and other alkaline ether extract the legal gas chromatography mass spectrometry, gas chromatography assay for quantification of chlorpromazine. 1.2 chlorpromazine the postmortem distribution studies in rabbits: rabbits six experiments before overnight fasting. Gavage uniform injection 8LD 50 (5990.4 mg / kg) of chlorpromazine. Be breathing and heartbeat disappeared rapidly anatomy of the animal, take the brain, heart, liver, spleen, lung, kidney, heart, cryopreservation, alkaline ether extraction, the legal gas chromatography mass spectrometry, gas chromatography assay for quantification of chlorpromazine. 2 dead after distribution research: male dogs 6 fixed on the dog on stage, gastric intubation, 2min injection 1/2LD 50 dose (201.6mg/kg according to body surface area conversion) chlorine promazine. 2h after hypoxia sacrificed supine position at room temperature (20 ° C), respectively, after the death of 0h, 2h, 4h, 8h, 12h, 24h, 48h, 72h, 96h fetch picking up material -20 ℃ refrigerator for inspection. Thawed, liquid - liquid extraction of blood and organs of the organizations, the legal nature of the gas chromatography mass spectrometry, gas chromatography assay for quantification of chlorpromazine. 3 after the death of diffusion study: 15 rabbits were randomly divided into five groups. Before overnight fasting, supine fixed rabbit Taiwan, gastric intubation, and then put to death; killed 1h after gavage uniform injection LD 50 (748.8mg/kg) chlorpromazine 3h, 1d, 2d, 4d and 6d each anatomical group (3) rabbit, take the brain, heart, liver, spleen, lung, kidney, left and right hindlimb muscle effort, peripheral blood, bile, urine, glass body fluids, -20 ℃ refrigerator for inspection. Thawed, liquid - liquid extraction of blood and organs of the organizations, the legal nature of the gas chromatography mass spectrometry, gas chromatography assay for quantification of chlorpromazine. Decomposition Kinetics Study: male dogs 6 fixed on the dog on stage, gastric intubation within 2 min injection LD 50 dose (403.2mg/kg) of chlorpromazine, administered volume of 20mL . 2h, after administration via the femoral artery was cannulated blood collection heparin; hypoxia were sacrificed, take efforts and liver. Each dog blood is divided into four equal parts were placed in 20 ° C, 4 ° C, -20 ° C, 20 ° C (plus NaF, 1%), each dog liver is divided into four equal parts were placed in 20 ° C, 4 ° C, -20 ℃, 20 ℃ (4% formaldehyde), which chlorpromazine # 0d, 7d, 54d, 90d, 150d, 280d, 320d gas chromatography mass spectrometry statutory sexual gas chromatography for quantitative detection. Using the the WinNonlin pharmacokinetic Software The average concentration of the drug treatment, fitting decomposition kinetics equation to calculate the decomposition half-life. 5. Statistical methods: using SPSS11.5 statistical software processing data, results are expressed as mean ± standard deviation ((x | -) ± S), t test. Results of gas chromatography and gas chromatography / mass spectrometry: GC / MS analysis of of Chlorpromazine the characteristic ions m / z 86,233,272,318. Chlorpromazine gas chromatography detection effort in regression equation, linear detection range, correlation coefficient, recovery rate, the minimum detectable concentration were Y = 4.2403X +0.2044 (μg / mL), 0.1 ~~ 10μg/mL 0.9927,99.03 ± 5.20%, 0.1μg/mL. Liver chlorpromazine gas chromatography detection regression equation, linear detection range, correlation coefficient, recovery rate, the lowest detectable concentration was Y = 10.38X +0.2784 (μg / mL), 0.1 ~~ 10μg/mL 0.9815,98.26 ± 2.50%, 0.1μg/mL. 2. Distribution of death: the 6LD 50 dose gavage to rats 10 ~ 11h death, chlorpromazine content in the body from high to low were: the stomach (2249.18 + / g) gt 164.23μg; liver ( GT; lung (571.09 ± 0.82μg / g) gt; brain (108.62 ± 27.38μg / g), spleen (105.37, ± of 17.64μg / g), kidney (100.33 ± 0.34μg / g and) 880.17 ± 8.62μg / g), heart (95.11 ± 4.63μg / g) gt; effort (6.31 ± 1.87μg/mL). 8LD 50 dose gavage rabbit 5 ~ 6h dead body chlorpromazine content from high to low were: stomach (3823.24 ± 662.66μg / g) gt; the Liver (489.67 ± 230.44μg / g ), lung (396.16 ± 237.70μg / g) gt; brain (251.24 ± 98.01μg / g), spleen (267.62 ± 87.46μg / g), kidney (the 289.01 ± 153.35μg / g), heart (117.88 ± 52.79μg / g) gt; effort (5.86 ± 1.90μg / mL). 3 dead after distribution: 1/2LD 50 chlorpromazine concentration first and then decreased in the gavage fatal dog effort tended to increase in concentration in the liver. Chlorpromazine content of the hard work in the postmortem stable 4h within 8 h was significantly higher (LSD, P <0.05), 48h ~ 96h within gradually reduce. Death and death effort chlorpromazine concentration than the (C postmortem blood / C death was blood << / sub> / sub> 0.47 to 2.28; chlorine in the liver the promazine content within 24h stabilize within 48h ~~ 96h was significantly higher (LSD, P <0.05), death and death was liver chlorpromazine concentration than the (C postmortem liver / C < sub> Death liver ) of 1.57 to 6.74; exposed dogs after the death of 96h, muscle chlorpromazine concentration the death no significant change (LSD, P> 0.05). 4 after the death of dispersion: after LD 50 chlorpromazine gavage rabbits 3h, were detected in the brain, heart, liver, spleen, lung, kidney, left and right hind limb muscle and effort chlorpromazine ingredients and spleen chlorpromazine highest concentrations in the urine and bile 6d after yet detected chlorpromazine ingredients. 5. decomposition kinetics: the decomposition kinetics of chlorpromazine in samples meet the first-order kinetics room open model, Ct = C 0 e -kt said 20 ° C, 4 ° C and stored at -20 ° C, 20 ° C (plus NaF, 1%) canine blood chloropropyl the triazine concentration 7d when dogs the chlorpromazine concentration decreased blood to the initial concentration of 57.6%, 97.3%, 53.6%, 80.2%, 20 ° C, 4 ° C, -20 ° C to 20 ° C (plus NaF, 1%) save 280d when canine blood concentration of chlorpromazine significant decline to the initial concentration of 29.80%, 36.51%, 19.62%, 39.29%, 20 ° C, 4 ° C, -20 ° C, 20 ° C (4% formaldehyde) to save the dog liver chlorine promazine concentration 7d rose to 237.01% of the initial concentration, 160.75%, 165.52%, 60.27%, save 320d, respectively, significantly decreased to 67.91% of the initial concentration, 78.60%, 67.84%, 12.63% Conclusion established of chlorpromazine in biological samples by gas chromatography / mass spectrometry and gas chromatography detection methods, the former good selectivity qualitatively accurate, the latter chromatography is simple, fast, agile, and quantitative results are accurate for chlorpromazine poisoning clinical rapid test diagnostic and chlorpromazine poisoning (dead) cases forensic identification. 2. chlorpromazine death unevenly distributed, 6LD 50 orally exposed rats chlorpromazine content order as : stomach, liver, lung, brain, spleen, kidney, heart, and hard work. 8LD 50 gavage rabbits chlorpromazine content in order for the stomach, liver, lung, brain, spleen, kidney heart, hard work. chlorpromazine concentration in the organs and tissues were higher than the blood in forensic identification of chlorpromazine the poisoning (dead) cases, in addition to the collection of blood and stomach, can also select the spleen, liver, lung, kidney, brain and other tissue samples, after the death of the distribution law of chlorpromazine poisoning cases forensic identification of the (dead) in the samples taken and special cases of blood drug concentration infer provide an experimental basis. 3. chlorpromazine in the exposed dogs can occur in vivo the Postmortem redistribution, 1/2LD 50 chlorpromazine in gavage death dogs death and death then effort chlorpromazine concentration ratio (C after death / C 0h ) of 0.47 to 2.28, after death and death was liver chlorpromazine concentration than (C after death / C 0h ) of $ 157 to 6.74, muscle content changed little. prompted chlorpromazine in beagle dogs exposed to death after the death of the time dependence of blood and organ concentrations can occur dead before distribution, the muscle tissue is less affected., chloropropyl The triazine accidental death or multiple drug abuse deaths forensic identification, not only to take the effort and try to acquisition muscle, brain and other distribution affect small samples to death before to consider postmortem time on the cause of death analysis, drawing The parts of corpses store environment and other factors, a comprehensive analysis. 4. chlorpromazine in the rabbits can occur after death dispersion, LD 50 chlorpromazine orally exposed to 3h, brain, heart, were detected in liver, spleen, lung, kidney, left and right hind limb muscle and effort the chlorpromazine ingredients, and spleen of chlorpromazine highest concentrations in the urine and bile in 6d yet detected chlorpromazine ingredients. the chlorpromazine hydrochloride poisoning (dead) Forensic Identification the lifetime taking poison and after the death of exposure can be detected in the urine and bile chlorpromazine to distinguish. 5. chlorpromazine the exposed dogs save the liver and efforts can degradation; storage temperature to reduce or add 1% of sodium fluoride can be extended so that the decomposition half-life and the detection period, and a 4% formaldehyde solution (20 ℃) ??fixed to be able to decompose to shorten the period of half-life and detection chloropropyl triazine decomposition kinetics in the sample compartment open model in line with the first-order kinetics, the available C t = C 0 e -kt said the decomposition kinetics equation, can be inferred anatomy or acquisition of the drug content in the samples was to provide an experimental basis for the toxicology test results of the analysis in the forensic identification of the poisoning cases (dead).
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