A rapid ultrasound-assisted dispersive liquid-liquid microextraction followed by ultra-performance liquid chromatography for the simultaneous determination of seven benzodiazepines in human plasma samples
Fernandez P.,Institute of Legal Medicine
Analytica Chimica Acta | Year: 2013
A simple and efficient ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) method has been developed for the determination of seven benzodiazepines (alprazolam, bromazepam, clonazepam, diazepam, lorazepam, lormetazepam and tetrazepam) in human plasma samples. Chloroform and methanol were used as extractant and disperser solvents, respectively. The influence of several variables (e.g., type and volume of dispersant and extraction solvents, pH, ultrasonic time and ionic strength) was carefully evaluated and optimized, using an asymmetric screening design 3242//16. Analysis of extracts was performed by ultra-performance liquid chromatography coupled with photodiode array detection (UPLC-PDA). Under the optimum conditions, two reversed-phases, Shield RP18 and C18 columns were successfully tested, obtaining good linearity in a range of 0.01-5 μg mL-1, with correlation coefficients r > 0.996. Quantification limits ranged between 4.3-13.2 ng mL-1 and 4.0-14.8 ng mL-1, were obtained for C18 and Shield RP18 columns, respectively. The optimized method exhibited a good precision level, with relative standard deviation values lower than 8%. The recoveries studied at two spiked levels, ranged from 71 to 102% for all considered compounds. The proposed method was successfully applied to the analysis of seven benzodiazepines in real human plasma samples. © 2013 Elsevier B.V.
Olze A.,Charité - Medical University of Berlin |
Solheim T.,University of Oslo |
Schulz R.,Institute of Legal Medicine |
Kupfer M.,Charité - Medical University of Berlin |
Schmeling A.,Institute of Legal Medicine
International Journal of Legal Medicine | Year: 2010
The question of whether an individual has reached the age of 18 is of crucial importance in forensic age estimation practice. In some countries, the age threshold of 21 years is relevant as well. A completed mineralization of third molars is not a sufficient criterion for a diagnosis of a minimum age of 18 years with the required probability. In a material of 1,198 orthopantomograms from 629 females and 569 males aged between 15 and 40 years, the visibility of the root pulp of fully mineralized lower third molars was evaluated according to stages 0, 1, 2, and 3. In females, stage 0 was first noticed at age 17.2 years, in males at age 17.6 years. In either sex, the earliest observation of stage 1 was between 21.0 and 22.4 years. Stage 2 was first achieved by males between 22.3 and 22.7 years, by females between 23.4 and 24.7 years. The occurrence of stage 3 was first found in both sexes between 25.1 and 25.9 years. These findings indicate that for stage 0, an age below 18 years cannot be excluded. However, for stage 1, the examined individual must be over 18 years of age and most probably over 21 years of age. For stages 2 and 3, the age can safely be stated to be over 21 years of age. This method may be a powerful tool for forensic dentists in age estimation in asylum and criminal proceedings. © Springer-Verlag 2010.
James Macaluso P.,Institute of Legal Medicine
Australian Journal of Forensic Sciences | Year: 2015
Only one study to date has explored the potential of innominate measurements for the estimation of stature in forensic contexts. The acetabulum, however, is among the skeletal regions most resistant to the processes of destruction and often survives intact. In this investigation, diameter, area and perimeter dimensions of the acetabulum were recorded from digital photographs in a documented French sample, consisting of 36 males and 33 females. Linear regression equations for estimating stature were then generated for each dimension in both males and females. Correlation coefficients for the models ranged from r = 0.401 to r = 0.474, and standard error of the estimate (SEE) values were between 5.7 cm and 6.1 cm. The highest correlation coefficients in both sexes were observed for the diameter of the acetabulum. These results indicate that acetabular dimensions should only be used for stature estimation when more accurate bones are unavailable for analysis. © 2014 Australian Academy of Forensic Sciences.
MacAluso Jr. P.J.,Institute of Legal Medicine |
Lucena J.,Institute of Legal Medicine
International Journal of Legal Medicine | Year: 2014
The ability to estimate stature from various parts of the body is essential in medicolegal investigations, particularly in cases in which human remains have been dismembered or mutilated. Therefore, the aim of this study was to generate linear regression models for predicting stature on the basis of sternum length in contemporary Spaniards. Sternal length, or the combined length of the manubrium and mesosternum, was recorded from digital radiographs of the chest plate for 65 males and 52 females. Correlation coefficients for the regression models were r∈=∈0.391 and r∈=∈0.461 for the male and female formulae, respectively. The standard errors of the estimate were 6.20 cm for the male equation and 5.56 cm for the female equation. Given these relatively large standard error rates, the derived stature estimation models should only be used when other more accurate skeletal elements, such as intact long bones, are unavailable for analysis. © 2014 Springer-Verlag Berlin Heidelberg.
Detection of genetic variation in KCNQ1 gene by high-resolution melting analysis in a prospective-based series of postmortem negative sudden death: Comparison of results obtained in fresh frozen and formalin-fixed paraffin-embedded tissues
Farrugia A.,Institute of Legal Medicine |
Keyser C.,Institute of Legal Medicine |
Ludes B.,Institute of Legal Medicine
International Journal of Legal Medicine | Year: 2012
High-resolution melting (HRM) analysis is a recently developed molecular technique proved to be applicable for detection of genetic variation, notably in sudden cardiac death. In certain circumstances, especially in postmortem genetic investigations, the formalin-fixed and paraffin-embedded (FFPE) tissues are the only DNA source available. The present study aimed to develop HRM assays, optimized for the analysis of FFPE tissues, to detect sequence variations in KCNQ1 exons in a prospective population-based series of postmortem negative sudden death and to compare the results between the paired freshly frozen and FFPE tissue samples simultaneously obtained from the same case. The analyses were conducted in each case of sudden death involving cases younger than 35 years with no significant morphological anomalies particularly with no cardiac structural disease and with negatives toxicological investigations. HRM analysis was successfully optimized for 13 of the 16 exons of the KCNQ1 gene. All mutated samples were correctly identified by HRM whatever the type of tissue tested. However, for FFPE samples, HRM indicated more positive samples than classical sequencing, used in parallel, due to the degradation of DNA by formalin fixation. This is the first postmortem study of KCNQ1 mutation detection with HRM on DNA extracted from FFPE samples with adapted protocol. Despite the false-positive detection, we concluded that the use of HRM as a screening method with FFPE samples to analyze KCNQ1 mutations can reduce the number of sequencing reactions and, thus, results in substantial time and cost savings. © 2012 Springer-Verlag.
Buttner A.,Institute of Legal Medicine
Handbook of Experimental Pharmacology | Year: 2010
Side effects of anabolic steroids with relevance in forensic medicine are mainly due to life-threatening health risks with potential fatal outcome and cases of uncertain limitations of criminal liability after steroid administration. Both problems are typically associated with long-term abuse and excessive overdose of anabolic steroids. Side effects may be due to direct genomic or nongenomic activities (myotrophic, hepatotoxic), can result from down-regulation of endogenous biosynthesis (antiandrogenic) or be indirect consequence of steroid biotransformation (estrogenic). Logically, there are no systematic clinical studies available and the number of causally determined fatalities is fairly limited. The following compilation reviews typical abundant observations in cases where nonnatural deaths (mostly liver failure and sudden cardiac death) were concurrent with steroid abuse. Moreover, frequent associations between structural characteristics and typical side effects are summarized. © 2009 Springer-Verlag Berlin Heidelberg.
Wulff B.,Institute of Legal Medicine
Forensic science international | Year: 2013
By German law organ and tissue donation may only be done if explicit consent is given during lifetime, e.g. by a donor card, or if the family grants informed consent after death in line with the deceased's last will, what is discussed to mean an additional emotional load in the situation of mourning. Every year about 35 musculoskeletal tissue donations and 250 donations of corneoscleral discs (means 125 donors) are performed in the Hamburg Institute of Legal Medicine. Talking to the next of kin plays a key role in the consenting process for donation. We show our guidelines according to the family contact by telephone and present the results of a catamnestic review referring to the donors of one year's period. Methods: One year after procurement had been completed we requested the consenting person to answer a questionnaire and asked for an evaluation of the contact to the institute's coordinator and furthermore the emotional impact of the consenting process. Results: 26 (72.2% of 36 only musculoskeletal donors) and 77 (73.3% of 114 only cornea donors) consenting persons answered and gave a positive feedback in the dimensions of information, support and consent. Discussion: The telephone contact with families of deceased persons out of the Institute of Legal Medicine is perceived to be helpful in an acute stress situation caused by a sudden death and an accepted way to establish the deceaseds' wishes in respect to donation through the relatives. Conclusion: According to the letter of the German Transplantation Act asking for donation is an obligation for every physician and our data show that this means no additional burden for relatives, if the question is placed appropriately. Forensic institutes play a key role in the recognition of tissue donors and take up the challenge to act as an interface between donors and recipients. But caring for the sufficient supply with tissue transplants needs considerable resources. Therefore additional support is necessary to fulfill this task. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.
Macaluso Jr. P.J.,Institute of Legal Medicine |
Lucena J.,Institute of Legal Medicine
International Journal of Legal Medicine | Year: 2014
A number of previous studies have demonstrated that osteometric analysis of the sternum provides a highly accurate method for discriminating adult sex in diverse population groups. In this study, sternal measurements were recorded from posteroanterior digital radiographs of the chest plate of 116 Spanish individuals (65 males and 51 females). Results demonstrated that all linear dimensions of the manubrium and mesosternum, sternal area, and sternal index were significantly sexually dimorphic in this population group. Discriminant function analyses incorporating several of these variables, individually or in combination, provided sex classification accuracy rates greater than 80.0 %, with associated sex biases below 5.0 %. A stepwise procedure, which can be used when a complete sternum is present, yielded the highest correct sex classification rate at 89.7 %. Only slightly lower allocation accuracy rates were obtained for multivariate equations which incorporated either dimensions of the manubrium or mesosternum (87.1 % for both formulae). Thus, the derived discriminant function equations should prove useful in forensic investigations, particularly those in which the pelvis or bones of the extremities are not available for analysis. © 2013 Springer-Verlag Berlin Heidelberg.
Cascallana J.L.,Institute of Legal Medicine |
Gordo V.,Institute of Legal Medicine |
Montes R.,Institute of Legal Medicine
Forensic Science International | Year: 2012
Methanol is a potent neurotoxic substance that causes severe metabolic acidosis and serious neurological disorders. Most of the cases are accidental exposures to drinking beverages contaminated with methanol. There are few articles reporting pure methanol intoxication; however, it is well known that small quantities of pure methanol causes blindness and death, the minimum lethal dose being 50-100. ml.A case report is presented of a 67-year-old woman, who committed suicide by ingestion of 500. ml of absolute methanol. Despite symptomatic and supportive intensive care, the woman died 23. h after hospital admission due to metabolic acidosis and multiple organ dysfunction syndrome. A complete medico-legal autopsy was performed. Grossly, there was complete detachment of the oesophagus mucosa and brownish discolouration of the gastric mucosa. Histological findings showed diffuse haemorrhagic necrosis of the stomach mucosa and intense acute inflammatory infiltration of the lamina propria. To our knowledge, this is the first autopsy report of such severe digestive injuries. A discussion and review of the recent literature on the subject are given. © 2012 Elsevier Ireland Ltd.
Sporkert F.,University of Geneva |
Kharbouche H.,University of Geneva |
Augsburger M.P.,University of Geneva |
Klemm C.,Institute of Legal Medicine |
Baumgartner M.R.,Institute of Legal Medicine
Forensic Science International | Year: 2012
In a case of a driving ability assessment, hair analysis for ethyl glucuronide (EtG) was requested by the authorities. The person concerned denied alcohol consumption and did not present any clinical sign of alcoholism. However, EtG was found in concentrations of up to 910. pg/mg in hair from different sampling dates suggesting an excessive drinking behavior. The person declared to use a hair lotion on a regularly base. To evaluate a possible effect of the hair lotion, prospective blood and urine controls as well as hair sampling of scalp and pubic hair were performed. The traditional clinical biomarkers of ethanol consumption, CDT and GGT, were inconspicuous in three blood samples taken. EtG was not detected in all collected urine samples. The hair lotion was transmitted to our laboratory. The ethanol concentration in this lotion was determined with 35. g/L. The EtG immunoassay gave a positive result indicating EtG, which could be confirmed by GC-MS/MS-NCI. In a follow-up experiment the lotion was applied to the hair of a volunteer over a period of six weeks. After this treatment, EtG could be measured in the hair at a concentration of 72. pg/mg suggesting chronic and excessive alcohol consumption. Overnight incubation of EtG free hair in the lotion yielded an EtG concentration of 140. pg/mg. In the present case, the positive EtG hair findings could be interpreted as the result of an EtG containing hair care product. To our knowledge, the existence of such a product has not yet been reported, and it is exceptionally unusual to find EtG in cosmetics. Therefore, external sources for hair contamination should always be taken into account when unusual cosmetic treatment is mentioned. In those cases, it is recommended to analyze the hair product for a possible contamination with EtG. The analysis of body hair can help to reveal problems occurring from cosmetic treatment of head hair. As a consequence, the assessment of drinking behavior should be based on more than one diagnostic parameter. © 2011 Elsevier Ireland Ltd.