Postmortem redistribution of fentanyl in the rabbit blood

Postmortem redistribution of fentanyl in the rabbit was investigated after application of the 50-μg/h Durogesic pain patch. Patches were applied for 48 hours. Two cycles of patch administration were used before characterization of the postmortem redistribution. Fentanyl showed marked redistribution into the femoral and pulmonary veins of the rabbit through 48 hours after the animals were humanely killed and the pain patches removed. The plasma concentration of 2.34 ng/mL in the femoral blood before killing the animals increased 5.6-fold by 48 hours after patch removal to 13.2 ng/mL. This postmortem concentration is approximately 3-fold the C(max) determined during antemortem pharmacokinetic analysis, 4 ng/mL, which was achieved 24 hours after the application of the second 50-μg/h Durogesic pain patch. After blood sampling for 48 hours after animal termination with patch removal compared with sampling for 48 hours from animals not terminated and with patch removal, the exposure ratios in the terminated animals were approximately 30-fold, indicating that between the postmortem redistribution of fentanyl and the cessation of hepatic clearance of fentanyl in the rabbit, the postmortem redistribution of fentanyl leads to an elevated measures of postmortem blood concentrations relative to antemortem blood concentrations.     

The extent of postmortem drug redistribution in a rat model.

The aim of this study was to investigate the postmortem redistribution of several drugs in a rat model and to examine if any of the pharmacological properties was related to the extent of this phenomenon. One of the following drugs: phenobarbital (phenobarbitone), acetaminophen (paracetamol), carbamazepine, codeine, verapamil, amphetamine, mianserin, trimeprazine (alimemazine) or chloroquine was administered together with nortriptyline orally to rats 90 min prior to sacrifice. Heart blood was sampled immediately before sacrifice and after 2 h postmortem, as it has previously been shown that this is sufficient time for postmortem concentration changes to occur in heart blood. Blood was also sampled from the clamped abdominal inferior vena cava (representing peripheral blood) and tissue samples were taken from lungs, myocardium, liver, kidney, thigh muscle, forebrain, and vitreous humor together with a specimen from the minced carcass. Drugs were analyzed by high performance liquid or gas chromatography. For phenobarbital, acetaminophen and carbamazepine the postmortem to antemortem blood drug concentration ratios were close to 1.0 and tissue concentrations were low. The postmortem to antemortem heart blood drug concentration ratio for chloroquine (6.9 +/- 1.5) was higher than for nortriptyline (3.5 +/- 0.3), and the remaining drugs (codeine, verapamil, amphetamine, mianserin, and trimeprazine) showed ratios of the same magnitude as nortriptyline. The postmortem to antemortem blood drug concentration ratios for both heart blood and blood from the vena cava and also the lung to antemortem blood drug concentration ratio were closely related to the apparent volume of distribution for the drugs studied (p < 0.001). Accordingly, an apparent volume of distribution of more than 3-4 L/kg is a good predictor that a drug is liable to undergo postmortem redistribution with significant increments in blood levels. The postmortem drug concentration in blood from vena cava was closely related to the antemortem blood level, confirming that among the postmortem samples, the peripheral blood sample was the most representative for the antemortem blood concentration.     

Timely antemortem and postmortem concentrations in a fatal carbamazepine overdose.

There are no published reports that include both timely antemortem and postmortem carbamazepine concentrations after massive overdose. We report a fatal overdose of carbamazepine with both timely antemortem and postmortem carbamazepine concentrations. Carbamazepine concentrations were 47.7 mcg/mL 2 h antemortem and 53 mcg/mL at 9 h postmortem. The slight rise in drug concentration may reflect continued absorption of the drug in the last 2 h before death. Postmortem carbamazepine concentrations drawn from a peripheral vessel in this patient appeared to reflect drug concentrations at the time of death.     

Drugs in postmortem adipose tissues: evidence of antemortem deposition

INTRODUCTION: Drug concentration measured in postmortem adipose tissue may or may not reflect antemortem concentration. To examine the possibility of whether the presence of basic drugs in adipose tissue is the result of postmortem change, we examined: tissues with and without livor mortis, concentration gradients within the adipose layer, and the stability of drug concentrations during the postmortem period. CASE REPORTS: Five drug-related deaths with case histories and analytical data are presented. Adipose tissues with and without livor mortis from the thigh area of the same decedent were analyzed for cocaine. The cocaine concentration of the tissue exhibiting 4+ livor was equivalent to the concentration observed in tissue without livor. Analyses of cross sections of adipose tissues containing cocaine and methamphetamine disclosed that drug concentrations were equally distributed throughout the layer, from just beneath the dermis to directly above the muscle. When morphine and temazepam concentrations were measured in adipose tissues collected from similar sites, but at different times, from the same cadaver, they remained essentially the same over 3 days (approximately 80 h). CONCLUSIONS: Since concentrations were the same in areas with and without livor mortis, the possibility of redistribution into adipose from blood or vascular channels is eliminated. The absence of a concentration gradient within the adipose layer rules out diffusion or permeation from muscle into the adipose layer, and the failure of morphine or temazepam concentration to change over time indicates that drugs in the adipose tissue are stable during the postmortem interval. Our findings support the notion that drugs identified in postmortem adipose tissue are there because of antemortem deposition and not because of any postmortem change or event.     

氯胺酮在急性中毒大鼠体内的死后再分布研究

目的探索氯胺酮在大鼠体内的死后再分布变化规律及温度对再分布的影响。方法48只雄性SD大鼠随机分为2个实验组（室温组24只、冷藏组18只）和1个对照组（6只）,实验组大鼠以氯胺酮290mg／kg灌胃,45min后缺氧处死,分别置于室温（24℃）和冷藏（4℃）条件下,于死后不同时间（0、12、24、48h）取心血、外周血、肝、肺、肾、心肌、大脑,检测其中氯胺酮含量;对照组大鼠以生理盐水灌胃,各对应组织器官样品为空白对照。血和组织样品中加入内标物SKF。。后碱化,乙酸乙酯萃取,GC／MS全扫描定性,内标法、工作曲线法气相色谱定量分析。结果室温条件下,大鼠死后48h内随着死亡时间延长,心血、肺、肝中氯胺酮的浓度呈升高趋势（P〈0．05）,肾脏中氯胺酮的浓度先升高后下降（P〈0．05）,外周血、心肌和脑中氯胺酮的浓度无显著性变化（P〉0．05）。冷藏条件下,血液及组织中氯胺酮浓度变化无显著性差异（P〉0．05）,除心肌外,各样本浓度均低于相应时段室温条件保存的样本。结论氯胺酮在大鼠体内存在死后再分布现象。温度对大鼠死后血液及组织中氯胺酮浓度变化有较明显的影响。     

Postmortem disposition of morphine in rats

The antemortem and postmortem distribution of morphine was studied in rats for the purpose of establishing whether drug distribution is altered after death. Samples were examined for free and total morphine concentration, pH and water content at 0-96 h after death. Morphine was administered antemortem at various intervals. All groups of rats studied showed a significant (P less than 0.05) increase in postmortem cardiac blood morphine concentrations. These changes, which are detectable within 5 min after death are likely to be related to an observed, rapid decrease in cardiac blood pH from 7.34 +/- 0.02 to 6.74 +/- 0.05. Significant increases in free morphine levels were, also, observed 24 and 96 h after death in liver, heart and forebrain while urine morphine levels decreased. The liver showed the greatest increase (20-fold) in free morphine levels 96 h after death, while hindbrain levels did not significantly change. Bacterial hydrolysis of morphine glucuronides accounted only in part for the observed increase in free morphine concentration. Postmortem fluid movement and pH-dependent drug partitioning was detected. It would appear that several mechanisms are responsible for postmortem drug distribution. Understanding the mechanisms and patterns responsible may eventually lead to better choices of postmortem tissue which may better represent antemortem drug levels.     

Avian vitreous humor concentrations of inosine, hypoxanthine, xanthine, uric acid, uracil and uridine as influenced by age and sex: their relevance as indicators of ante-mortem hypoxia

An investigation was made to determine the effects of age and sex on postmortem concentrations (mumol/l) of inosine, hypoxanthine, xanthine, uric acid, uracil and uridine in the vitreous humor of chickens (Gallus domesticus). Five male and 5 female chickens were sampled each week from 0-10 weeks of age. Samples were collected at 0 and 24 h postmortem and analysed by HPLC. Hypoxanthine, uric acid and uridine were detected at both 0 and 24 h postmortem whereas inosine, xanthine and uracil were detected only at 24 h postmortem. Neither sex nor the sex X age interaction influenced the concentrations of the compounds investigated (P greater than 0.01). Age had a significant influence on the concentrations of all compounds at 24 h postmortem (P less than 0.0001). At 0 h postmortem, age significantly affected the levels of hypoxanthine and uridine (P less than 0.0001) but not uric acid (P = 0.014). All compounds occurred at higher concentrations at 24 than 0 h postmortem with the exception of uridine at 0 and 1 week of age. Previous studies using data confounded by age and postmortem sampling time have concluded that postmortem vitreous humor hypoxanthine concentration is a useful indicator of antemortem hypoxia. The results of this study cast doubt on that conclusion. Age and postmortem sampling time should be rigorously controlled when conducting quantitative analyses of these compounds in vitreous humor.     

Evaluation of the Postmortem Glucose and Glycogen Levels in Hepatic,Renal, Muscle,and Brain Tissues: Is It Possible to Estimate Postmortem Interval Using These Parameters?

The aim of our study was to investigate the postmortem levels of glucose and glycogen in hepatic, renal, muscle, and brain tissues and then examine the changes in those levels that could be useful for estimating postmortem interval. We established an animal model. Seventy female BALB/c albino mice were used in this study. After being sacrificed, the mice were randomly divided into six groups according to time elapsed since death (Group 1: 0 h; Group 2: 12 h; Group 3: 24 h; Group 4: 36 h; Group 5: 48 h; and Group 6: 60 h). Glucose levels were significantly different between groups for all tissues studied. Slope of the change per unit time was higher for the hepatic glucose levels. Based on these results, it is possible to estimate postmortem interval using postmortem glucose levels in hepatic tissue. Tissue‐specific assessment may contribute valuable information to postmortem interval studies.     

氯氮平在家兔体内死后再分布规律研究

目的阐明死后48h内家兔体内氯氮平再分布规律,为相关法医鉴定工作提供借鉴。方法取家兔15只,随机分为5组,以氯氮平灌胃,分别于死后0、6、12、24、48h取心血、外周静脉血、尿液、肝组织检测氯氮平浓度。结果家兔死亡后心血、外周静脉血、肝脏氯氮平浓度不断升高,尿液氯氮平浓度不断降低;死后早期浓度变化率大于晚期浓度变化率。死后48h心血、外周静脉血、肝脏、尿液氯氮平浓度分别为死后0h各检材氯氮平浓度的418%、193%、154%和29%。结论死亡一段时间后,提取生物检材,检测出的氯氮平浓度并不能准确反映刚死时的实际浓度。     

Quantitation of nucleotides,nucleosides and bases in antemortem and postmortem bloodstains by high-performance liquid chromatography

Ante- and post-mortem bloodstains prepared from the blood of volunteers and corpses were analysed for ATP and its related compounds by reversed-phase high-performance liquid chromatography (HPLC). The results showed that (1) ATP was present in a large amount in antemortem bloodstains but not in postmortem stains, (2) AMP, adenosine, inosine, hypoxanthine, xanthine and uracil either were not detected or were detected in smaller amounts in antemortem than in postmortem bloodstains, and (3) ADP was present in both ante- and postmortem bloodstains. These differences suggest that quantitation of these compounds may be useful in identifying whether bloodstains are ante- or post-mortem.     

不同温度下离体人静脉血ATP含量与死亡时间的关系

目的探讨在环境温度变化条件下,人体静脉血ATP降解与死亡时间的关系。方法健康志愿者48名,随机分为6组,肱静脉取静脉血,置于10℃、15℃、20℃、25℃、30℃和35℃下保存;每4h应用ATP检测仪对不同温度下的ATP含量进行检测;应用SPSS统计软件进行回归分析,MATLAB软件进行差值函数分析拟合。结果各温度组ATP值随死亡时间延长均呈下降趋势,从取血即刻的1 573.683 E-13mol/L,降至6.00 E-13mol/L左右,所经历的时间分别为236h(10℃)、163h(15℃)、124h(20℃)、92h(25℃)、72h(30℃)和64h(35℃),对所得数据进行回归分析,得到各温度组下ATP含量变化与PMI关系的二元三次曲线方程(R2范围为0.976～0.990);进行差值拟合,得到10～35℃范围内ATP含量变化与PMI关系的三元四次曲面方程。结论在不同温度下,人体静脉血ATP降解与PMI关系符合三元四次方程分布,利用差值函数拟合的方法可在温度变化条件下进行死亡时间推断。     

Linearization of the relationship between postmortem plasma chloride concentration and postmortem interval in rats

A potentiometric titration procedure was employed for measurement of plasma chloride concentrations during the 0-96 h postmortem period in rats. The data revealed antemortem absolute values and postmortem rate of decrease in plasma chloride concentration (PCl) which were almost identical to those in dogs and in man. Expression of the data in the form of a double logarithmic plot of PCl versus postmortem interval (PMI) yielded a linear relationship of high correlation (r = -0.97; P less than 0.001). Attention is drawn to the possibility of utilizing postmortem rate of change in plasma chloride concentration as a means of estimating time elapsed since death.     

Complement component C3a or C3a desArg as a new marker for estimation of local vital reactions in incised skin wounds.

The anaphylatoxin C3a or its desArg form (C3a/desArg) generated during complement activation could be detected in the vicinity of incised skin wounds of guinea pigs using immunoblotting methods. The C3a/desArg peptides were detectable immediately after injury in local sites up to 3 mm from the wound edge. In subsequent determinations of up to at least 3-day-old antemortem wounds, the maximum concentration of these peptides was largely localized up to 6 mm from the wound edge at 2 h after injury. In postmortem wounds, however, these peptides were undetectable. When they were released in antemortem wounded tissues they could be detected up to 1 day at 22 degrees C after death. These results suggest that the detection of C3a/desArg in wounds using immunoblotting methods can be useful for distinguishing ante- from postmortem wounds.     

大鼠皮肤切创后E-选择素表达及稳定性研究

目的探讨大鼠皮肤切创后E-选择素表达规律及法医学意义。方法健康SD大鼠90只,随机分成4组：正常对照组、活体切创组（30min～7d12个时间点）、死后切创组（30min～3h3个时间点）、死后稳定性组（-20％6h-7d9个时间点,25％6h～3d5个时间点）。在大鼠头部建立皮肤切创模型,按设定的时间点取皮肤检材,运用免疫组化和图像分析技术,检测血管内皮E-选择素的表达规律。结果在活体切创组中,伤后1hE-选择素在血管内皮细胞内即呈阳性表达,持续至伤后7d,且随时间变化呈规律性表达。在正常对照及死后切创组未见阳性表达。死后-20％稳定性组各时间点E-选择素表达与死后即刻比较无显著性差异（P〉0．05）。25％各时间点与死后即刻比较有显著性差异（P〈0．01）。结论E-选择素在创伤后血管内皮细胞内特异性的表达具有时序性规律,且低温条件下稳定性较好。     

Assessment of the Role Played by N‐propanol Found in Postmortem Blood in the Discrimination Between Antemortem Consumption and Postmortem Formation of Ethanol Using Rats

This study disproves the reliability of n‐propanol as a biomarker to establish whether the ethanol found in postmortem blood is derived from antemortem ingestion or postmortem putrefactive processes. Two groups of rats were given ethanol or normal saline solution, respectively, and sacrificed 1.5 h later. After putrefaction, blood and, in a few cases, urine samples from the rats were analyzed for ethanol and n‐propanol by head‐space gas chromatography equipped with flame ionization detection. Although the concentration ratios of ethanol/n‐propanol in the postmortem blood collected from the bodies without prior alcohol consumption were expected to be <20 (as per limited case reports and previous in vitro studies), in samples from several rats that were on saline solution, this ratio was found to exceed 20. In conclusion, the concentration ratio of ethanol/n‐propanol in postmortem blood does not allow for the discernment between antemortem ingestion and the postmortem synthesis of ethanol.     

大鼠肾细胞核 DNA含量与死亡时间关系的研究

目的 研究 DNA含量与死亡时间关系。方法 应用计算机图像分析技术,测定了 15只大鼠在死后 48h内不同时间点大鼠肾细胞核 DNA含量的面积（ Area）、等效直径（ Mean－ Dia, MD）、异形指数（ Index of density, ID）、平均光度（ Average optical density, AOD）、积分光度（ Integral optical density, IOD）、密度变化数（ L－ Den－ Coe, LDC）和平均灰度（ Average gray, AG）等七项参数的变化值。结果 在大鼠死亡的相对早期（ 48h内）,其细胞核 DNA降解速率与 PMI具有一定相关性。本实验并将每个参数的测量值进行了多项式运算,获得了更能体现 DNA降解趋势的二项式回归方程。结论 应用计算机图像分析技术,测量机体死后 DNA含量变化,将会成为推断死亡时间精确、客观的新方法。     

尸体心血红细胞ATP降解规律与早期死亡时间的相关性

目的观察早期死亡时间（PMI）与血液红细胞ATP含量的相关性。方法选择具有确切死亡时间的尸体30例,在死亡后6、8、10、12、14、16、18、20、22、24h分别于第4肋间进行心脏穿刺取血,利用生物发光法检测血液样本红细胞ATP含量（μmol／gHb）,并观察红细胞ATP含量变化与死亡时间的关系。结果尸体心血红细胞ATP含量在死亡后1～24h之内呈现非匀速下降趋势,与死亡时间的Pearson相关系数为-0．971（P=0．000）;尸体心血红细胞ATP含量与死亡时间的回归方程及尺。值为：Y=-0．096X＋2．872（x为死亡时间）,R2=0．936,P=0．000。结论尸体心血红细胞ATP含量在死后1—24h之内的变化与死亡时间具有相关关系,可以作为法医学死亡时间推断的生物学指标。     

实验性大鼠急性心肌缺血的酶组织化学观察

本文报告50只雄性Wistar大鼠结扎左冠状动脉所致早期急性心肌梗死（缺血1～8h的组织化学变化。结果发现：糖原含量在缺血1h、NADHD和LDH活性在缺血2h梗死区明显下降；缺血4h，酶活性下降发展到全部缺血区。CCo.CK及ATP酶活性下降在缺血4h明显，缺8h发展到整个缺血区。非缺血区NADHD、LDH、CK、ATP酶活性能耐受死后自溶的影响达24h以上，具有法医学实践意义。糖元（PAS染色）和PPr在死后早期降解快，非缺血区与缺血区无显著性差异，因此缺乏法医病理学应用意义。     

References for determining the time of death by potassium in vitreous humor

The different statements concerning the slope and intercept of the regression line and the 95% limits of confidence are the reason that potassium in vitreous humor is not used (at least in Germany) as an aid in estimating the time of death. The relationship between the concentration of potassium and the time of death is mainly influenced by antemortem electrolyte imbalances caused by disease and/or duration of terminal episode. The influence of terminal episode is best identified by its duration (Adelson et al., J. Forensic Sci., 8 (1963) 503-514). In order to have a method suitable for every case and to be as precise as possible we looked therefore for parameters in vitreous humor which were stable postmortem and indicating antemortem electrolyte imbalance. Urea is such a parameter, being stable postmortem (Coe, Am. J. Clin. Pathol, 51 (1969) 741-750) and useful as a marker of antemortem electrolyte imbalance. Our investigations on potassium in vitreous humor, including sudden and hospital deaths after chronic lingering disease, revealed 95% limits of confidence of +/- 34 h up to 120 h postmortem. Reviewing only cases with urea less than 100 mg/dl the 95% limits of confidence could be reduced to +/- 22 h. Considering the duration of terminal episode (less than 6 h) the precision was +/- 20 h. In this way our modified procedure is suitable for every case with the resulting precision of estimation being determined only by the duration of the terminal episode and urea concentration.     

电击死兔骨骼肌及心肌HSP70 mRNA和c-fos mRNA表达

目的 研究电击死兔骨骼肌与心肌HSP70 mRNA和c-fos mRNA-表达变化。探究生前电击与死后电击的鉴别方法。方法 15只新西兰兔,随机分电击死组、死后电击组和对照组,每组5只,用荧光RT-PCR技术检测骨骼肌与心肌热休克蛋白70（HSP70） mRNA与c-fos mRNA表达水平,对所得结果进行统计学分析。结果 生前电击兔骨骼肌及心肌HSP70 mRNA与c-fos mRNA表达高于死后即刻电击者（P〈0．05）。结论 检测骨骼肌及心肌HSP70 mRNA与c-fos mRNA表达变化有助于于生前电击与死后电击的鉴别。