| Objective:1. To establish the simultaneous determination of 8 psychotherapeutic agents in samples.2. To establish an amitriptyline acute poisoning model of rats, study the distribution of amitriptyline in acute poisoning rats. Obtain amitriptyline distribution characteristics and regularity in rats.3. Take amitriptyline for example, to study the stability of psychotherapeutic agents in fluids and tissues of poisoning rats in different storage temperatures.Methods1. Gas Chromatography (GC) methods: After adding SKF525A (internal standard), body fluids and tissues were acidified and extracted by diethyl ether. Qualitative analysis was based on retention time. Quantitative analysis was based on internal standard method and working curves method.2. Distribution study: The rats in experimental group were given intragastric administration of amitriptyline with a dose of 392mg/kg and executed by cervical dislocation. Blood, urine, heart, liver, lung, kidney, muscle and brain were taken out to detect amitriptyline. Rats in control group were given physiologic saline. The samples were taken out correspondingly and used as blank check.3. Stability study: the rats in experimental group were given intragastric administration of amitriptyline with a dose of tow times of LD50 and executed by cervical dislocation. Blood, lung, liver were preserved at various temperatures (4℃, -20℃) to detect amitriptyline at different time points. Rats in control group were given intragastric administration of physiologic saline. The samples were taken out correspondingly and used as blank check. Results:1. The peak of 8 psychotherapeutic agents is successively Miltown, Amitriptyline, Doxepin, Diazepam, Penfluridol, Clozapine, Haloperidol and Chlorprpmazine. The linear range of amitriptyline in blood, urine and liver are 5.0～200.0μg/mL, 5.0～200.0μg/mL, 1.0～150.0μg/g respectively. The limit of detection in blood, urine and liver are 1.0μg/mL,1.0μg/mL,0.5μg/g respectively. Coefficients of recoveries in blood of different concentration are 105.6±1.5%, 96.6±4.3% and 97.5±3.1%. Coefficients of recoveries in urine of different concentration are 105.2±3.4%, 100.8±3.6% and 95.5±3.9%.2. The concentration of amitriptyline in biological tissues and fluids of acute poisoned rats was as follows: lung﹥liver﹥spleen, muscle, kidney﹥brain﹥heart﹥blood﹥urine.3. The concentration of amitriptyline changed in biological tissues and fluids of acute poisoned rats at various temperatures (4℃, -20℃): The concentration of amitriptyline in lung and liver were stable at -20℃. The concentration of amitriptyline in lung and liver were 125.54% and 117.07% of the initial concentration up to 70 days. Along with the increase of saved time the concentration of amitriptyline increased significantly at -20℃. The concentration of amitriptyline in blood, lung and liver samples changed greatly at 4℃. In blood, the trend was decline - rose– downward. In lung and liver, the trends were decline– up. With the increase of freezing time, impurity peak in lung and liver increased to interfere with the detection of amitriptyline, the concentration can not be quantified accurately.Conclusion:1. The analytical method could determine several psychotherapeutic agents simultaneously. This method is of nice accuracy, high sensitivity and handle easily. It is suitable for forensic toxicological analysis and iatrogenic poisoning determination.2. Distribution in rats those were given intragastric administration of amitriptyline showed: The concentration of amitriptyline in lung, liver and spleen is high with the highest concentration in lung. Besides blood and stomach content, lung, liver and spleen could be selective specimens for determination of amitriptyline in case of amitriptyline poisoning.3. The temperature and storage time affected amitriptyline content in different samples. The concentration of amitriptyline in lung and liver were stable at -20℃. The concentration of amitriptyline in different samples changed greatly at 4℃and impurity peak disturbed the detection. 4℃was unfavorable conditions for keeping samples. For blood samples, it is better to detect them as soon as possible.
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