Determination of opiates in postmortem bone and bone marrow.

Bone and bone marrow of a fatally poisoned heroin addict were analyzed by FPIA and GC-FID, immediately after death. A piece of the bone from the above case was buried for 1 year and analyzed by the same procedure. Morphine was detected in all specimens at concentrations of 195, 340 and 155 ng/g for bone marrow, bone and buried bone, respectively. A loss of 54.4% of morphine concentration was observed during 1-year burial. Such findings have potential forensic value in cases of skeletonized remains.     

Methamphetamine and amphetamine concentrations in postmortem rabbit tissues

The feasibility of detecting methamphetamine and its major metabolite, amphetamine, in postmortem tissues over a 2-year period was examined. It is important to determine if the abuse and toxic effects of drugs can be proved from evidence found in decayed, submerged, or stained tissue materials. The blood, urine, liver, skeletal muscle, skin and extremity bones from rabbits given methamphetamine intravenously were kept at room temperature, under 4 different conditions: sealed in a test tube, dried in the open air, submerged in tap water and stained on gauze. Methamphetamine was present in all the samples, with slight change in concentration in case of sealed and air dried tissues. Changes varied in bones kept in water. There were considerable decreases in methamphetamine in blood and urine stains. Despite long term storage, drug abuse and/or toxicity could be determined, in all tissues examined.     

Site-dependent postmortem changes in blood cocaine concentrations

When a forensic toxicologist interprets postmortem blood cocaine findings he usually must make assumptions regarding perimortem drug concentrations. In-vitro studies have shown that cocaine rapidly hydrolyzes in unpreserved blood, particularly at elevated temperatures. However, other studies have demonstrated site-dependent postmortem release of some drugs from tissue stores accompanied by increases in drug concentrations in the blood. This study was undertaken to investigate whether blood cocaine concentrations change in the body during the postmortem interval and, if so, to measure the direction and magnitude of the changes. In medical examiner cases in which scene investigation suggested that the decreased was a cocaine user, blood samples were collected as soon after death as possible. At autopsy, a second set of samples was collected. Analysis of paired samples by gas chromatography/mass spectrometry (GC/MS) revealed dramatic differences in the cocaine concentration. The magnitude and direction of the change appears to be site dependent. Usually, but not invariably, cocaine concentration in subclavian vein blood decreases while that in heart, aorta, and femoral vein blood increases during the interval between death and autopsy. The findings emphasize the danger inherent in attempting to estimate the concentration of cocaine in blood at the time of death from postmortem data.     

离体人胸骨骨髓DNA降解与腐败尸体死亡时间推断的初步研究

目的研究死后胸骨骨髓DNA的降解与较长死后间隔时间(PMI)的关系。方法采用改良Feulgen染色及计算机图像分析技术对离体人胸骨(取材后分别室温搁置0,1,3,5,7d)骨髓DNA含量进行定量检测。结果在较长PMI范围内,人胸骨骨髓DNA含量仍呈现下降规律。结论人胸骨骨髓DNA降解规律可望应用于较长PMI的推断。     

Codeine and morphine blood concentrations increase during blood loss

During extensive blood loss, a plasma volume refill will take place by transfer of extravascular fluid into the circulation. Drugs present in this fluid may follow and cause a rise or a drop in blood drug concentration, depending on their levels and accessibility in the restoration fluid. This study explored the possible changes of codeine, and its metabolite morphine, in whole blood during a standardized exsanguination in the rat. Three doses containing 5 mg codeine were given orally. In eight rats, blood loss was accomplished by slowly withdrawing 0.8 mL blood at 10 min intervals during 70 min. In control rats, blood was withdrawn only at 0 and 70 min. At 70 min, the final/initial codeine and morphine concentration ratios were 0.70 +/- 0.38 and 0.88 +/- 0.47, respectively, in controls, but increased to 1.28 +/- 0.44 (p = 0.014) and 1.41 +/- 0.34 (p = 0.021), respectively, in exsanguinated rats. It is concluded that blood loss can affect blood drug concentrations.     

The influence of site of collection on postmortem morphine concentrations in heroin overdose victims

When assaying for postmortem morphine concentration, significant site sampling variability exists between central and peripheral sampling sites and even within sampling regions of the body. To study the variation, 76 suspected heroin overdoses were identified. Each had femoral artery (FA) and vein (FV), left and right ventricle and pooled heart blood samples obtained at autopsy. Forty-four tested positive for morphine. Morphine concentrations were determined by gas chromatography/mass spectrometry, with sampling site differences reported as log-transformed ratios and compared by signed rank test.The mean FA to FV ratio for total morphine was 1.2 (range 0-4.5). The ratio for left heart to right heart total morphine was 1.1 (range 0.4-3.2). Left ventricular to FV total morphine ratio was 2.0 (range 0.6-6.9). In these opioid overdose deaths, FA and FV morphine concentrations are usually similar, although up to 4.5-fold differences were noted. Centrally obtained morphine concentrations are on average twice as high compared with peripheral morphine concentrations. Left and right ventricular morphine concentrations were usually similar, although up to 3.2-fold differences were noted (left side higher).     

Blood versus bone marrow pentobarbital concentrations

Postmortem pentobarbital levels in rabbit heart blood and bone marrow were determined and compared. The average ratio of femur marrow/blood pentobarbital concentrations in 24 rabbits was 1.06 +/- 0.05. The average percent difference between actual plasma pentobarbital concentrations and calculated plasma pentobarbital concentrations was 5.82 +/- 1.96. Concentrations were determined by gas chromatography of extracted, derivatized pentobarbital.     

Blood versus bone marrow isopropanol concentrations in rabbits

The use of bone marrow to determine the blood isopropanol concentrations becomes important when a blood specimen is contaminated or unavailable. The blood/marrow isopropanol ratios were determined in rabbits autopsied 0, 4, and 24 h after sacrifice. The lipid content of the individual marrow specimens was shown to have a significant influence on the range of ratios. When the determined marrow isopropanol concentrations were corrected for lipid content, a better correlation between blood and marrow concentrations was obtained. The ratio (1.45 ± 0.17) was not altered significantly by postmortem time or temperature.Although acetone was not exogenously administered to the rabbits, but rather was endogenously produced from isopropanol metabolism, the relationship between blood and marrow acetone concentrations was somewhat linear. However, the range of observed and corrected blood/marrow acetone ratios was altered significantly by storage temperature, and delays between death and analysis. Thus, under the experimental conditions of this study, marrow isopropanol concentrations may be used to predict blood isopropanol concentrations, whereas marrow acetone concentrations can not.     

Blood versus bone marrow methanol concentrations in rabbits

Postmortem methanol levels in bone marrow and heart blood were determined in rabbits. The average ratio of heart blood concentration to observed bone marrow concentration in 36 rabbits was 2.6 ± 0.6 with a range of 1.5 to 4.2. Correcting for the lipid content of the bone marrow decreased the average ratio, reduced the ratio range and improved the correlation. The heart blood to corrected bone marrow ratio was 1.6 ± 0.3 with a range of 1.2 to 2.9. Direct injection gas chromatographic techniques were employed to quantitate methanol concentrations.     

Some observations concerning blood morphine concentrations in narcotic addicts

Blood samples from deceased narcotic addicts were analyzed for morphine, and the results form persons who died from narcotic addiction were compared with those from homicide victims. In most instances morphine was detectable in both types of death, and usually the values obtained were less than 30 microgram/dl. Narcotic addiction deaths involving only morphine, or morphine plus a combination of ethanol, quinine, or diazepam (Valium), were also evaluated. In some cases high quantities of ethanol were present, and death could be attributed to the combined CNS depressant effects of morphine and ethanol. The quinine levels would not normally be considered toxic, however, and it could not be ascertained that the quantity of this drug present contributed to death. Diazepam was present in elevated concentrations, and its depressant effect may have been a factor in some narcotic addiction deaths.     

A comparison of carboxyhemoglobin saturation values in postmortem heart blood and peripheral blood specimens

The following is a study conducted to determine whether there was any significant difference in carboxyhemoglobin (COHb) saturation levels between the heart blood and blood collected from a peripheral site. The average heart blood to peripheral blood COHb saturation level ratio in the 42 cases studied was 1.09. Sixty-two percent (26 of 42) of the cases had a heart blood to peripheral blood ratio between 0.9 and 1.1; 74% (31 of 42) had a ratio between 0.8 and 1.2. Eighty-three percent (35 of 42) had a ratio between 0.7 and 1.3. There were four cases where the heart blood to peripheral blood ratio was either below 0.6 or greater than 1.4. The differences between the two sites were not statistically significant.     

Postmortem blood free and total morphine concentrations in medical examiner cases

This purpose of this study was to determine the relationships between postmortem free morphine and total morphine levels in a large series of medical examiner morphine and heroin related deaths. Free morphine, total morphine, and 6-monoacetylmorphine (6-MAM) concentrations were measured by gas chromatography-mass spectrometry (GC-MS) in 87 medical examiner cases over 20 months. The mean total morphine concentration, mean free morphine concentration, and mean percent free morphine for all cases were: 2.3 mg/L (SD 5.2 mg/L), 0.5 mg/L (SD 1.6 mg/L), and 19.4% (SD 22.8%); respectively. Regression analyses showed weak correlations between total and free morphine concentrations over the entire concentration range (0 to 36.6 m/L, r = 0.603, n = 91) and over a subset concentration range of 0 to 1.0 mg/L (r = 0.369, n = 54). Twenty-three out of 56 (41%) tested positive for 6-MAM, indicative heroin abuse cases. Lower total and free morphine concentrations and a higher percent free morphine were found in individuals with detectable 6-MAM. Comparing blood concentrations for cases with and without detectable 6-MAM demonstrated mean total morphine concentrations of 0.9 mg/L versus 2.1 mg/L (p = 0.05), mean free morphine concentrations of 0.3 mg/L versus 0.4 mg/L (p = 0.21), and mean percent free morphine of 34.7% versus 13.7% (p < 0.003), respectively. Our findings demonstrate higher free to total morphine ratios in individuals with detectable 6-MAM than in individuals without 6-MAM. The database established in this study may assist medical examiners in the evaluation of postmortem blood opiates regarding the cause of death in opiate related ingestion cases.     

Fatal versus non-fatal heroin "overdose": blood morphine concentrations with fatal outcome in comparison to those of intoxicated drivers

The study was performed to distinguish fatal from non-fatal blood concentrations of morphine. For this purpose, blood levels of free morphine and total morphine (free morphine plus morphine conjugates) in 207 cases of heroin-related deaths were compared to those in 27 drivers surviving opiate intoxication. The majority of both survivors and non-survivors were found to show a concomitant use of depressants including alcohol or stimulants. Blood morphine levels in both groups varied widely, with a large area of overlap between survivors (free morphine: 0-128 ng/ml, total morphine: 10-2,110 ng/ml) and non-survivors (free morphine: 0-2,800 ng/ml, total morphine: 33-5,000 ng/ml). Five (18.5%) survivors and 87 (42.0%) non-survivors exhibit intoxication only by morphine. In these cases, too, both groups overlapped (survivors-free morphine: 28-93 ng/ml, total morphine: 230-1,451 ng/ml; non-survivors-free morphine: 0-2,800 ng/ml, total morphine: 119-4,660 ng/ml). Although the blood levels of free or total morphine do not allow a reliable prediction of survival versus non-survival, the ratio of free/total morphine may be a criterion to distinguish lethal versus survived intoxication. The mean of the ratio of free to total morphine for all lethal cases (N=207) was 0.293, for those that survived (N=27) 0.135, in cases of intoxication only by morphine 0.250 (N=87) and 0.080 (N=5), respectively. Applying a cut-off of 0.12 for free/total morphine and performing ROC analyses, fatal outcome can be predicted in 80% of the cases correctly, whereas 16% of the survivors were classified as dead. Nevertheless, in this study, all cases with a blood concentration of 200 ng/ml and more of free morphine displayed a fatal outcome.     

Blood Versus bone marrow ethanol concentrations in rabbits and humans

Post-mortem ethanol levels in bone marrow and heart blood were determined in rabbits and humans. The average ratio of blood/femur marrow ethanol in 39 rabbits was 2.10 ± 0.23 with a range of 1.27 – 3.50. In 18 human autopsy cases the average ratio of blood/rib marrow ethanol levels was 2.16 ± 0.32 with a range of 1.65 – 2.94. The average blood/marrow ratios in humans and rabbits were comparable. However, a wider range of ratios was noticed in the rabbit study.     

A storage study of ethanol in rabbit and human bone marrow

The study reports the effect of storage on ethanol concentrations in bone marrow.     

Sulfide concentrations in postmortem mammalian tissues

Postmortem changes in sulfide concentrations in body tissues were examined in autopsied rats exposed to hydrogen sulfide concentrations of 550 to 650 ppm, and in nonexposed rats and humans. Analyses were made by gas chromatography, following an extractive alkylation. Sulfide concentrations in the blood, liver, and kidneys of rats increased in both the exposed and nonexposed groups, depending on the lapse of time after death. On the other hand, the lung, brain, and muscle showed little or no change in sulfide concentration with elapse of time after death. The data obtained from human tissues were almost the same as those for rats, except data for blood, in which no or little increase of sulfide was observed.