| Objectives1. To explore the best extraction separation method, establishment of ketamine in rat tissues in gas chromatography-mass spectrometry (Gas Chromatography/Mass Spectrometry, GC/MS) quantitative analysis method.2. Using the established method of ketamine in main organs, low, high dose of ketamine for anatomy of different time and harvested after death in rat heart, liver, spleen, lung and kidney were measured in. The experimental data were analyzed to clarify, low, high dose of ketamine to ketamine main organs of drug in rat postmortem redistribution, of low, high dose of ketamine administration influenced the distribution of ketamine in major organs of rats after death.3. Using the established method, the ethanol with low, high dose of ketamine combined administration of ketamine in main organs and were sacrificed at different time after anatomy materials in rat heart, liver, spleen, lung and kidney were measured in. The experimental data were analyzed, clarifying ethanol with low, high dose of ketamine combined administration of ketamine in major organs of rats after death and distribution, to investigate the effect of ethanol on the low, high dose of ketamine to major organs in rats of ketamine drug postmortem redistribution.Methods1. Determination of ketamine and ketamine combined with GC/MS method of ketamine in main organs of ethanol in rat heart, liver, spleen, lung and kidney. In the1.00G organ homogenates adding internal standard propyl adiphenine (Proadifen hydrochloride, SKF-525A)50μg,2times of extraction at pH13with ethyl acetate3min shaking after centrifugation. The organic phase with, in80℃water bath and drying, the residue with100μL chromatographic pure methanol volume,1uL sample analysis. By comparing retention time qualitative, quantitative internal standard working curve method. 2. Grouped:according to the random number table,60healthy male Sprague-Dawley rats were divided into ketamine injection group (n=30) and ketamine combined with alcohol group (n=30). Ketamine injection group by ketamine dosage were divided into low dose group and high dose group, low dose group and high dose group and press were anatomy were divided into low dose group (based, killed immediately after n=5), low dose group were sacrificed48h after (here in after referred to as the low dose group48h, n=5) death, low dose72h after group (here in after referred to as the low dose72h group, n=5) and control group (high dose were killed immediately after, n=5), high dose group were sacrificed48h after (hereinafter referred to as the high dose48h group, n=5), high dose group were sacrificed72h after (hereinafter referred to high dose72h group, n=5). Ketamine in ethanol group by ketamine dosage were divided into low dose group and high dose combined with ethanol ethanol group, low dose group and high dose combined with ethanol ethanol group and press were anatomy were divided into low dose ethanol control group (combination of materials, killed immediately after n=5), low dose of ethanol were used48h based group (hereinafter referred to as the low dose combined with48h group, n=5), low dose combined with ethanol were72h based group (here in after referred to as the low dose combined with72h group, n=5) and control group (high dose and ethanol were killed immediately after, n=5), high dose combined with ethanol were killed48h after high dose group (here in after referred to as in group48h, n=5), high dose combined with ethanol were72h based group (hereinafter referred to as the high dose combined with group72h, n=5). The number of each animal were. According to low dose ketamine group (30mg/kg) and high dose (60mg/kg) by intraperitoneal injection of ketamine administration. Ketamine in ethanol group by2mL/kg dose administered5min with20%ethanol induced by low dose (30mg/kg) and high dose (60mg/kg) by intraperitoneal injection of ketamine administration. The rats were continuously administered for30days (1times, each time interval of24h) and at15min after the last administration. All rats in control the body organ distribution map, according to the grouping of anatomy, separation of the target organs and tissues, after treatment, according to the GC/MS analysis method to detect the content of ketamine, and uses the SPSS software to analysis of statistical analysis.Results1. Methods of qualitative and quantitative analysis The organization of ketamine, GC/MS analysis method, the concentration in the range of20-1000μg/ml good linear relationship, the correlation coefficient γ>0.9997.The lowest detection of ketamine limit was10μg/ml, the lowest detection limit is5ng/ml, the recovery rate of>85.0%, precision RSD<3.99%days, between day precision<5.5.2. The main organs of ketamine injection of ketamine in rats postmortem redistribution 2.1Low dose (30mg/kg)group:Ketamine content in the heart, compared with the low dose group, low dose48h group showed an upward trend, there is significant difference (p<0.01); low dose72h group decreased, but no significant difference. Ketamine content in the liver, compared with the low dose group, low dose48h group and low dose72h group decreased with increased, there was no significant difference in. Ketamine content in spleen, low dose48h group than in the low dose group increased, and there is a significant difference (p<0.01); low dose72h group, low dose48h group decreased, but still higher than the low dose group. Ketamine content in the lungs, compared to the low dose group, low dose48h group and low dose group72h showed a downward trend, which has significant difference between low dose72h group (p<0.05); ketamine content in kidney, compared with the low dose group, low dose48h group and low dose72h group were decreased trend, and there was a significant difference (p<0.01).2.2High dose (60mg/kg) groups:Ketamine content in the heart, compared with the high dose group, high dose48h group and high dose group72h ketamine increased, and there was a significant difference (p<0.01). Ketamine content in the liver, compared to the high dose group, high dose48h group and high dose72h group were increased, and there was a significant difference (p<0.05, p<0.01). Ketamine in the spleen, compared with the high dose group, high dose48h group and high dose72h group showed a downward trend, which has significant difference between the group of high dosage of72h (p<0.01). Ketamine content in the lungs, compared to the high dose group, high dose48h group and high dose group72h ketamine content declined, and there were significant differences (p<0.05). Ketamine content in kidney, compared to the high dose group, high dose48h group and high dose72h group showed a downward trend, which has significant difference between the group of high dosage of72h (p<0.05).2.3Low dose group and high dose group:Ketamine content in the heart, high dose group, high dose48h group, high dose of ketamine in72h group than those in low dose group increased, but no significant difference. Ketamine content in the liver, high dose group, high dose48h group, high dose of ketamine in72h group than those in low dose group increased, there were significant differences between48h in high dose group, low dose48h group and high dose72h group and low dose72h group (p<0.05). Ketamine in spleen in the high dose group, low dose group increased, and there is a significant difference (p<0.01); high dose48h group, high dose72h group compared with the low dose48h group, low dose72h group were decreased, but no significant difference. Ketamine content in the lungs, high dose group, high dose48h group, high dose72h group than those in low dose group increased, which had significant difference between high dose72h group compared with the low dose of72h (p<0.05). Ketamine content in the kidney, the high dose group and control group at low concentration, high dose48h group, high dose of ketamine in72h group than those in low dose group increased, but no significant difference. 3The posthumous redistributions of ketamine in the main organs of rats to ketamine combined with alcohol group3.1Low dose ketamine combined with ethanol group:the content in the heart, combined with low dose of ethanol compared to the control group, low dose group and low dose combined with48h72h group increased, and there was a significant difference (p<0.01). Ketamine content in the liver, compared with the low dose combined with ethanol control group, low dose group and low dose of48h combined with72h group showed an upward trend, but no significant difference. Ketamine in the spleen, compared with the low dose combined with ethanol control group, low dose combination of48h group increased, and there is a significant difference (p<0.05); low dose combined with low dose72h group with48h group decreased, but still higher than that of low dose combined with ethanol control group, and there was significant difference (p<0.05). Ketamine content in the lungs, compared to low doses combined ethanol control group, low dose combination of48h group increased, and there is a significant difference (p<0.01); low dose combination of72h group was higher than that of low dose combined with ethanol control group, compared to the low doses combined48h group decreased, and there were significant difference (p<0.01). Ketamine content in kidney, low dose combination of48h group was higher than that of low dose combined with ethanol control group, and there were significant difference (p<0.01); low dose combined with low dose72h group with48h group decreased, but still higher than that of low dose combined with ethanol control group, and there were significant differences (p<0.01).3.2High dose combined with ethanol group:Ketamine content in the heart, the high dose combined with48h group and high dose72h group with high dose combined with ethanol control group was essentially flat, no significant difference. Ketamine content in the liver, high dose48h group with high dose combined with ethanol decreased, and there were significant difference (p<0.01); high dose plus72h group, higher dose in48h group increased, but still lower than the high dose combined with ethanol control group, and there were significant difference (p<0.01). Ketamine content in spleen, high dose48h group with high dose combined with ethanol decreased, and there were significant difference (p<0.01); high dose plus72h group, higher dose in48h group increased, but still lower than the low and high dose combined with ethanol control group, with significant difference (p<0.0l). Ketamine content in the lungs, high dose48h group with high dose combined with ethanol control group showed an upward trend, and there is a significant difference (p<0.01); high dose plus72h group, higher dose in48h group decreased, there was significant difference (p<0.01), and lower than the high dose combined with ethanol control group; ketamine content in kidney the combination of48h, high dose group and high dose group were higher doses combined with72h ethanol control group showed an upward trend, but no significant difference. 3.3Comparing low dose group with high dose of ethanol in ethanol groups:Ketamine content in the heart, with high dose ethanol control group, high dose group, high dose of48h combined with72h group were better than those of the corresponding low dose group increased, the combination of high dose ethanol control group were significantly different between the ethanol control group combined with low dose (p<0.01). Ketamine content in the liver, high doses combined ethanol control group, high dose group, high dose of48h combined with72h group than in the low dose group increased firstly and then decreased, the content of the high dose combined with ethanol compared to the control group in low doses combined ethanol control group increased, and there is a significant difference (p<0.01); ketamine content in spleen, high doses combined ethanol control group, high dose group, high dose of48h combined with72h group than in the low dose group increased firstly and then decreased, the combination of high dose ethanol control group than those in the low dose combined with ethanol control group, there were significant differences (p<0.01). Ketamine content in the lungs, the high dose combined with ethanol control group, high dose group, high dose of48h combined with72h group than in the low dose group increased, the combination of high dose ethanol control group compared to the low doses combined ethanol control group, there were significant differences (p<0.05). Ketamine content in kidney, high doses combined ethanol control group, high dose group, high dose of48h combined with72h group compared with the low dose group increased firstly and then steady downward trend, but no significant difference.4Effects of ethanol on posthumous redistribution of ketamine in the main organs of ketamine dependence rats4.1Low dose group and low dose group compared with ethanol:Ketamine content in the heart, low doses combined ethanol control group, low dose48h group than those in the low dose group decreased, the low doses combined alcohol control group and low dose group had significant difference (p<0.01) and low dose combination;72h group was higher than the group of low dosage of72h increased slightly, but no significant difference. Ketamine content in the liver, low doses combined ethanol control group, low dose group, low dose of48h combined with72h group than in the low dose group showed the first decline after rising trend, but no significant difference. Ketamine content in spleen, low doses combined ethanol control group, low dose group, low dose of48h combined with72h group than in the low dose group showed increasing trend, in which low doses combined alcohol control group and low dose group had significant difference (p<0.05). Ketamine content in the lungs, low doses combined ethanol control group, low dose group, low dose of48h combined with72h group than in the low dose group showed increasing trend after the first drop, in which low doses combined alcohol control group and low dose group had significant difference (p<0.01); ketamine content in kidney, low doses combined ethanol control group, low dose group, low dose of48h combined with72h group than in the low dose group showed increasing trend after the first drop, in which low doses combined alcohol control group and low dose group had significant difference (p<0.01).4.2High dose groups and high dose of ketamine in combination:content of heart, high doses combined ethanol control group, high dose group, high dose of48h combined with72h group than in the high dose group increased at first and then decreased, the combination of high dose ethanol control group than in control group with high dose significant differences (p<0.01). Ketamine content in the liver, high doses combined ethanol control group, high dose group, high dose of48h combined with72h group than in the high dose group increased at first and then decreased, and there was a significant difference (including high doses combined ethanol control p<0.01control group, between the null distribution group and high dose and high dose combination between48h group and high dose48h group and high dose combination of p<0.05between72h group and high dose72h). Ketamine content in spleen, high doses combined ethanol control group, high dose group, high dose of48h combined with72h group than in the high dose group increased at first and then decreased, with the high dose combined with ethanol control group than in the high dose group had significant difference (p<0.05). Ketamine content in the lungs, the high dose combined with ethanol control group, high dose group, high dose of48h combined with72h group than in the high dose group decreased and then increased and then decreased, with the high dose combined with ethanol control group than in the high dose group had significant difference (p<0.05); the content of ketamine in kidney high dose ethanol, with control group, high dose48h group than in the corresponding combination of high dose group decreased, the combination of high dose ethanol control group than in the high dose group had significant difference was significant difference (p<0.01).Conclusion:1. A high sensitivity, high selectivity, high recovery rate, simple and efficent quantitative analysis method for ketamine in related viscera rat has been Established2. Injection doses of ketamine can affect posthumous redistributions of ketamine in rat heart, liver, spleen, lung, kidney to a considerable degree. As the contents of ketamine In heart, liver and kidney of ketamine injection combined with ethanol group show a downward trend compared with those of ketamine injection group, the contents of ketamine in spleen and lung are not much changed, either increased or decreased. This change may be caused by the promoting metabolism of ethanol. |
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