Study On The Forensic Toxicokinetics Of Chlorpromazine

Objective1. To establish the analysis of the chlorpromazine in living samples by a GC and a GC/MS.2. To develop a postmortem distribution,postmortem redistribution, postmortem diffusion anddecomposition kinetics model of chlorpromazine.3. Study the postmortem distribution in rats and rabbits,postmortem diffusion in rabbits,postmortem redistribution and decomposition kinetics of chlorpromazine in dogs,and provide ascientific evidence for the forensic identification of chlorpromazine poisoning death.Methods1. The postmortem distribution of chlorpromazine:1.1 The postmortem distribution of chlorpromazine in rats:6 male SD rats, weight 200g±10gwere given chlorpromazine through stomach with a dose of 1350 mg/kg. As soon as the vitalsigns disappeared,the rats were dissected and the specimens such as heart blood, heart, liver,spleen, lung, kidney, brain were sampled immediately, in which the concentration ofchlorpromazine was determined by a GC and a GC/MS.1.2 The postmortem distribution of chlorpromazine in rabbits: 6 rabbits were givenchlorpromazine through stomach with a dose of 5990.4mg/kg.As soon as the vital signsdisappeared,the rabbits were dissected and the specimens such as heart blood,heart,liver,spleen,lung,kidney,brain, were sampled immediately, in which the concentration ofchlorpromazine was determined by a GC and a GC/MS.2. The postmortem redistribution of chlorpromazine: 6 male dogs were given chlorpromazinethrough stomach with a dose of 201.6 mg/kg was administered, the dogs were put to death for 2hours after administration, then placed at 20℃, the heart-blood, liver, muscle was taken atdifferent time point of 0h, 2h, 4h, 8h, 12h, 24h, 48h, 72h, 96h after the death respectively,qualitative and quantitative analysis was performed by GC/MS and a GC equipped with a NPD.3. The postmortem diffusion of chlorpromazine:15 rabbits were randomly allocated to fivegroups. After being executed by an aeroembolism for 1 hour, 15 rabbits were given a dose of748.8mg/kg through stomach.Three animals were randomly dissected after postmortemadministration of chlorpromazine at 3h, 1d, 2d, 4d, 6d respectively and the specimen such asbrain, heart, liver, spleen, lung, kidney, muscle of right and left lower limb,heart blood, blood,bile, urine, vitreous fluid were sampled and stored at -20℃in refrigerator for detection,qualitative and quantitative analysis was performed by a GC/MS and a GC equipped with NPD.4. The decomposition kinetics of chlorpromazine: 6 male dogs were given chlorpromazinethrough stomach with a dose of 403.2 mg/kg was administered, the dogs were put to death after administration. The heart-blood were collected and divided equally to four group in 20℃, 4℃,-20℃and 20℃(add NaF to 1%)and the liver were collected and divided equally to four group in20℃,4℃,-20℃and 20℃(4% methanal)were taken at different time point of 0h, 7d, 54d, 90d,150d, 280d, 320d after the death respectively, qualitative and quantitative analysis wasperformed by a GC/MS and a GC equipped with NPD. The equation and sparameters ofdecomposition kinetics were imitated and calculated with Winolin program.5. Statistics method:The datas was collected by software SPSS11.5 and expressed by((x|-)±S).Results1. GC/MS and GC:The m/z of characteristic ion of chlorpromazine was 86, 233, 272, 318 by aGC/MS.The linear range of chlorpromazine in biomaterial by a GC was 0.1 to 10μg/mL. Theregression equation, linear range,correlation coefficient, recovery, limit of detection ofphenobarbital in heart blood was Y=2.5464X-0.2919(μg/mL), 0.1～10μg/mL, 0.9927,96.43±5.20ï¼…, 0.1μg/mL. The regression equation, linear range, correlation coefficient, recovery,limit of detection of chlorpromazine in liver was Y=10.38X＋0.2784(μg/mL), 0.1～10μg/mL,0.9815, 98.26±2.50ï¼…, 0.1μg/mL.2. Postmortem distribution: After the administration of chlorpromazine, the 6LD₅₀ group ratsdied after the administration for 10h to 11h and the 8LD₅₀ rabbits died for 5h to 6h. The order ofphenobarbial detected in rats was as follows:(1) 6LD₅₀ group:stomach (2249.18±164.23μg/g)>live(880.17±8.62μg/g)>lung (571.09±0.82μg/g) >brain(108.62±27.38μg/g), splee(105.37±17.64μg/g), kidney (100.33±0.34μg/g), heart(95.11±4.63μg/g)>heart blood(6.31±1.87μg/ml) (2)8LD₅₀ stomach (3823.24±662.66μg/g)> liver(489.67±230.44μg/g), lung(396.16±237.70μg/g) > brain(251.24±98.01μg/g), splee (267.62±87.46μg/g), kidney(289.01±153.35μg/g), heart(117.88±52.79μg/g)> heart blood (5.86±1.90μg/g).3. Postmortem redistribution:The1/2LD₅₀ dogs died after the administration , the chlorpromazineconcentrations detected in the heart-blood present a first rise then decrease trend, in the liverpresent a rise trend. The chlorpromazine content in heart blood within 4 hour after the death isstability, then significantly higher at 8 hour (LSD,P<0.05)and gradually reduce at 48h～96h. Thechlorpromazine concentration ratio between postmortem and at the death reach 0.47～2.28. Thechlorpromazine content in liver within 24 hour after the death is stability, then significantlyhigher within 48h～96h hour (LSD, P <0.05), The ratio of chlorpromazine concentrationbetween postmortem and at the death reach 1.57～6.74; The chlorpromazine concentrationdetected in the muscle of died dogs at 96 hour after the death is no significantly change (LSD, P>0.05).4. Postmortem diffusion:Expect for bile and urine,chlorpromazine was detected in all otherspecimens sampled at 3h after the postmortem administration of chlorpromazine in rabbits. The concentration of chlorpromazine in spleen reached a peak value. And the chlorpromazine stillcan not be detected in urine and bile at 6^th d after the death.5. Decomposition kinetics:The decomposition kinetics of chlorpromazine were fit to the towcompartment open model with a first order kinetics.The common equation was C_t=C₀e^-kt .Concentrations in the blood of the dogs were 57.6%, 97.3%, 53.6%, 80.2% of the initial analysesdecreased to 7 days at 20℃, -20℃, 20℃(add NaF to 1%). Concentrations in the blood of thedogs were 29.80%, 36.51%, 19.62%, 39.29% of the initial analyses decreased to 280 days at 20℃, 4℃, -20℃, 20℃(add NaF to 1%). Concentrations in the liver of the dogs were237.01%,160.75%, 165.52%, 60.27% of the initial analyses up to 7 days at 20℃,4℃,-20℃,20℃(4%methanal). Concentrations in the liver of the dogs were 67.91%, 78.60%, 67.84%, 12.63% of theinitial analyses decreased to 320d at 20℃, 4℃, -20℃, 20℃(4% methanal).Conclusion1. The GC/MS and GC method is more selective, more sensitive, simpler, more accurate, and canbe applied not only to the clinical diagnosis of chlorpromazine poisoning, but also to forensicidentification of chlorpromazine poisoning death.2.The order of chlorpromazine contents detected in poisoning death rats was stomach, live, lung,brain, splee, kidney, heart, heart blood. The order of chlorpromazine contents detected inpoisoning death rabbitts was stomach, liver, lung, brain, splee, kidney, heart, heart blood. Theconcentration of chlorpromazine in blood was lower than in other organs. So, besides the bloodand stomach, more organs should be collected spleen, liver, lung, kindey, brain and postmortemdistribution provide a scientific evidence for the forensic identification of chlorpromazinepoisoning death.3. Chlorpromazine exibited postmortem redistribution in poisoning death dogs. The1/2LD₅₀ dogsdied after the administration, the chlorpromazine concentration ratio between postmortem and atthe death in heartblood reach 0.47～2.28. The chlorpromazine concentration ratio betweenpostmortem and at the death in liver reach 1.57～6.74. The chlorpromazine concentrationdetected in the muscle of died dogs after the death is no significantly change. It was suggest thatthe chlorpromazine in dead dogs exist the blood of postmortem time-relay and the change oforgans concentration.4. There was a postmortem diffusion of chlorpromazine in rabbits. Expect for bile and urine,chlorpromazine was detected in all other specimens sampled at 3h after the postmortemadministration of chlorpromazine in rabbits. The concentration of chlorpromazine in spleen reacha peak value. And the chlorpromazine still can not be detected at 6d. In chlorpromazinepoisoning (death) forensic identify, the chlorpromazine in urine and bile whether be detected ornot to distinguish poison after death and poisonous. 5. The decomposition kinetics of chlorpromazine were fit to the first order kinetics. The commonequation was Ct=C₀e^-kt and k was the decomposition constant of first order reaction, which couldused to conclude the initial concentrations of chlorpromazine in corpse and to provide ascientific evidence for the forensic identification of the anesthesia accident caused bychlorpromazine.