International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s004140050012

Issue: Volume 113, Number 2

Date: February 2000

Pages: 0117 - 0120

Demonstration of morphine in ganglion cells of the hippocampus from victims of heroin overdose by means of anti-morphine antiserum

F. Wehner, H.-D. Wehner, J. Subke, R. Meyermann, P. Fritz

^A1 Institut für Gerichtliche Medizin der Universität Tübingen, Nägelestraße 5, D-72074 Tübingen, Germany Tel. +49-7071-2972031; Fax +49-7071-551732
^A2 Institut für Hirnforschung der Universität Tübingen, Calwerstraße 3, D-72076 Tübingen, Germany Tel. +49-7071-2982283
^A3 Institut für Pathologie am Robert Bosch Krankenhaus, Auerbachstraße 110, D-70376 Stuttgart, Germany Tel. +49-711-81013394

Abstract:

Abstract To investigate the topography of morphine distribution in the human brain, a method has been developed to detect morphine immunohistochemically. In this study hippocampus tissue from victims of heroin overdose (blood morphine concentrations 220 ng/g-1500 ng/g; 6-MAM positive urine sample), known for its high concentration of w-opiate receptors was used. The immunohistochemical staining was performed with an anti-morphine antiserum originally developed for radio-immuno-assays. In comparison with control specimens from cases of sudden death without morphine exposition or a history of heroin abuse, the brains from victims of heroin overdose showed selectively stained ganglion cells, axons and dendrites, suggesting a massive concentration of morphine in the neuronal structures.

Keywords:

Key words Ganglion cells · Hippocampus · Immunohistochemistry · Mean optical density (MOD) · Morphine

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s004140050073

Issue: Volume 110, Number 4

Date: June 1997

Pages: 226 - 229

Antibody-mediated clean-up of blood for simultaneous HPLC determination of morphine and morphine glucuronides

J. Beike ^A1, H. Köhler ^A1, G. Blaschke ^A2

^A1 Institut für Rechtsmedizin, Westfälische Wilhelms-Universität, von Esmarchstrasse 86, D-48149 Münster, Germany Fax: +49 (251) 8355 158
^A2 Institut für Pharmazeutiche Chemie, Westfälische Wilhelms-Universität, Hittorfstrasse 58, D-48149 Münster, Germany

Abstract:

Abstract For the interpretation of the concentration of morphine in blood samples of heroin consumers information about the concentration of the analgesic active morphine metabolite morphine-6-glucuronide is very important. Thus a simple but specific clean-up procedure based on immuno-affinity chromatography is presented for the extraction of morphine, morphine-3-glucuronide and morphine-6-glucuronide from whole blood in cases of fatal heroin overdose. The preparation of the immunoabsorber by immobilization of antibodies against morphine-3-BSA and morphine-6-KLH with carbonyldiimidazole-activated trisacrylgel is described. The separation of the extracts is achieved by HPLC using native fluorescence detection. The limits of detection for this method are 10 ng for morphine and morphine glucuronides/g blood. The results for the concentration of morphine and morphine glucuronides in blood from seven cases of heroin overdose are presented. By calculating the quotients for the concentrations of morphine-6-glucuronide/morphine the time elapsed since the last intake of heroin is estimated.

Keywords:

Key words Morphine glucuronides · Antibody-mediated · clean-up · Immunoabsorber · HPLC native fluorescence · detection

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s004140050201

Issue: Volume 112, Number 1

Date: December 1998

Pages: 62 - 66

Fatal injections of heroin. Interpretation of toxicological findings in multiple specimens

H. Druid ^A1, P. Holmgren ^A2

^A1 Division of Forensic Medicine, Faculty of Health Sciences, University of Linköping, S-581 85 Linköping, Sweden Fax: +46 13 101 182
^A2 Division of Forensic Chemistry, Faculty of Health Sciences, University of Linköping, S-581 85 Linköping, Sweden

Abstract:

Abstract We report two fatalities due to injection of heroin. The first case was witnessed but during the early phase of the police investigation the question was raised whether the injection was self-administered. Multiple samples were collected from different sites and analysed in order to establish drug distribution and to determine the site of injection. Fresh injection marks were found in both antecubital fossae but histological examination failed to settle which one was the last. However, toxicological analysis of the tissues at the injection sites indicated that the injection in the right arm was the last one. This was consistent with the suspicion that the victim was given the injection by another person although probably in agreement with the deceased. In the second case, a similar toxicological procedure was used. This fatality was not witnessed, however ample evidence indicated that it was an isolated event in a former intravenous heroin addict and there was only one fresh injection mark. Even in this case, the concentration of morphine was much higher in the tissue sample from the injection mark than in any of the blood samples.

Keywords:

Key words Heroin · Morphine · Fatal · Blood-concentration · Disposition · Injection

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

Issue: Volume 113, Number 2

Date: February 2000

Pages: 0102 - 0106

Effect of the shampoo Ultra Clean on drug concentrations in human hair

J. Röhrich, S. Zörntlein, L. Pötsch, G. Skopp, J. Becker

^A1 Institut für Rechtsmedizin, Johannes Gutenberg-Universität Mainz, Am Pulverturm 3, D-55131 Mainz, Germany
^A2 Institut für Rechtsmedizin und Verkehrsmedizin, Ruprecht-Karls-Universität Heidelberg, Voßstrasse 2, D-69115 Heidelberg, Germany

Abstract:

Abstract The influence of the special shampoo Ultra Clean (Zydot Unlimited, Tulsa, Oklahoma) on the results of hair analyses was investigated. Hair samples from persons (n = 14) with a known history of drug abuse were collected at autopsy. The hair samples were divided into separate strands which were analyzed both after washing with Ultra Clean and without treatment. Hair analyses were performed by methanol extraction under sonication, purification by solid phase extraction and GC/MS in SIM mode according to routine procedures for tetrahydrocannabinol (THC), cocaine, amphetamine, methylenedioxyamphetamine (MDA), methylenedioxymethamphetamine (MDMA), methylenedioxyethylamphetamine (MDE), heroin, 6-monoacetylmorphine (6-MAM), morphine, codeine, dihydrocodeine and methadone. All drugs originally present in the hair fibers were still detected after a single ¶application of Ultra Clean. However, a slight decrease in drug concentrations could mostly be observed e.g. cocaine (n = 10) -5%, 6-MAM (n = 12) -9%, morphine (n = 12) -26%, THC (n = 4) -36%. The findings clearly demonstrated that drug substances had not been sufficiently removed from human hair by a single Ultra Clean treatment to drop their concentrations below the limit of detection of the analytical method applied.

Keywords:

Key words Hair · Drug testing Hair analysis · Manipulations · Shampoo effect

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

Issue: Volume 113, Number 3

Date: May 2000

Pages: 150 - 154

On-site testing of saliva and sweat with Drugwipe and determination of concentrations of drugs of abuse in saliva, plasma and urine of suspected users

N. Samyn ^A1, C. van Haeren ^A1

^A1 Nationaal Instituut voor Criminalistiek en Criminologie, Vilvoordsesteenweg 100, B-1120 Brussels, Belgium e-mail: walter.samyn@rug.ac.be, Tel. +32-2-2400500, Fax +32-2-2400501 or +32-2-2416105

Abstract:

Abstract Potential drug users participated voluntarily in a Belgian study on the usefulness of the non-instrumental immunoassay Drugwipe (Securetec, Germany) for the screening of cocaine, opiates, amphetamine and cannabinoids in saliva and sweat. If one of the screening assays (urine, oral fluid, sweat) showed a positive result, blood and saliva were collected. The on-site Drugwipe results were correlated with the Drugwipe results for saliva in the laboratory and with the GC/MS results of the corresponding saliva, plasma and urine samples and pharmacological effects at the time of sampling. The Drugwipe assay proved to be sufficiently sensitive for the detection of recent cocaine (n = 6) and amphetamine (n = 15) abuse, whether the device was wiped on the tongue or on the surface of the body, or when a saliva sample was applied to the wiping part. In five of the six potential cocaine users, the saliva concentrations of cocaine exceeded 1000 ng/ml. In the amphetamine group, the saliva concentrations of amphetamine, MDMA or both were high (> 1000 ng/ml) in 13 subjects. For cocaine and amphetamine, the positive scores for Drugwipe matched the GC/MS results for the three body fluids. Recent heroin abuse (n = 5) could be demonstrated to some extent with Drugwipe on samples from the tongue but only the two subjects with the highest saliva concentrations of MAM (> 500 ng/ml) and morphine (> 500 ng/ml) were positive. If the legal cut-off value for driving under the influence of opiates in Belgium (20 ng/ml of free morphine in plasma) was taken into account, only three subjects would have been legally positive. For cannabinoids (n = 15), false negatives and even some false positives were observed. Saliva can be considered as a useful analytical matrix for the detection of drugs of abuse after recent abuse when analysed with GC/MS.

Keywords:

Key words Saliva · Drugs of abuse · Drugwipe · GC/MS · Driving under the influence

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s004140050190

Issue: Volume 112, Number 1

Date: December 1998

Pages: 8 - 14

A specific immunoassay for the determination of morphine and its glucuronides in human blood

J. Beike ^A1, G. Blaschke ^A2, A. Mertz ^A3, H. Köhler ^A1, B. Brinkmann ^A1

^A1 Institut für Rechtsmedizin, Westfälische Wilhelms-Universität, Von-Esmarch-Strasse 62, D-48149 Münster, Germany
^A2 Institut für Pharmazeutische Chemie, Westfälische Wilhelms-Universität, Hittorfstrasse 58, D-48149 Münster, Germany
^A3 Alkamed SARL, 2, avenue de l’Energie, F-67800 Bischheim, France

Abstract:

Abstract The development of specific antisera for immunochemical determination of morphine, morphine-3-glucuronide and morphine-6-glucuronide is described. Morphine was N-demethylated to normorphine and N-alkylated to give N-aminopropyl-normorphine as hapten for antisera against morphine. As haptens for antisera against morphine-3-glucuronide and morphine-6-glucuronide, N-aminopropyl-nor-morphine was glucuronidated in position 3 or 6 respectively. Each of these three haptens were coupled to BSA employing the glutaraldehyde method to obtain three different immunogens. Immunisation of rabbits with these conjugates gave anti-morphine, anti-morphine-3-glucuronide and anti-morphine-6-glucuronide antisera, which were tested in a competitive, heterogeneous radioimmunoassay. Tracers for this radioimmunoassay procedure were synthesised by substitution of morphine and morphine-6-glucuronide in position 2 with ¹²⁵I and indirect iodination of the morphine-3-glucuronide hapten according to the method of Bolton and Hunter. The resulting antisera show very specific reactions with morphine, morphine-3-glucuronide and morphine-6-glucuronide. Cross reactivities of each antiserum with structurally related opiates and opioides are very low. The cross reactivities of the anti-morphine antiserum against morphine-3-glucuronide, morphine-6-glucuronide, codeine, codeine-6-glucuronide or dihydrocodeine were less than 0.3%, the anti-morphine-3-glucuronide antiserum against morphine, morphine-6-glucuronide, codeine, codeine-6-glucuronide or dihydrocodeine less than 0.1% and the anti-morphine-6-glucuronide antiserum against morphine, morphine-3-glucuronide, codeine or dihydrocodeine less than 0.1%, against codeine-6-glucuronide less than 2.3%. The determination of morphine, morphine-3-glucuronide and morphine-6-glucuronide in blood samples (limit of detection = 3, 1, 0.5 ng/g) of nine cases of fatal heroin overdose with this radioimmunoassay method and the comparison with a GC/MS method is described.

Keywords:

Key words Morphine · Morphine-3-glucuronide · Morphine-6-glucuronide · Radioimmunoassay · RIA · GC/MS

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s00414-002-0290-2

Issue: Volume 117, Number 1

Date: February 2003

Pages: 2 - 9

Immunohistochemical demonstration of the amphetamine derivatives 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) in human post-mortem brain tissues and the pituitary gland

Els A. De Letter, Marc F. A. Espeel, Marijke E. C. Craeymeersch, Willy E. Lambert, Karine M. Clauwaert, Riet Dams, Kjell A. Mortier, Michel H. A. Piette

^A1 Ghent University, Department of Forensic Medicine, J. Kluyskensstraat 29, 9000, Ghent, Belgium
^A2 Ghent University, Department of Anatomy, Embryology, Histology and Medical Physics, Godshuizenlaan 4, 9000, Ghent, Belgium
^A3 Ghent University, Department of Toxicology, Harelbekestraat 72, 9000, Ghent, Belgium
^A4 Ghent University, Department of Medical Biochemistry and Clinical Analysis, Harelbekestraat 72, 9000, Ghent, Belgium

Abstract:

Abuse of amphetamine derivatives such as 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) is an important issue in current forensic practice and fatalities are not infrequent. Therefore, we investigated an immunohistochemical method to detect the amphetamine analogues MDMA and MDA in human tissues. For the staining procedure, the Catalysed Signal Amplification (CSA) method using peroxidase (HRP) provided by Dako^® and specific monoclonal antibodies were used. Appropriate controls for validation of the technique were included. The distribution of these designer drugs was studied in various brain regions including the four lobes, the basal ganglia, hypothalamus, hippocampus, corpus callosum, medulla oblongata, pons, cerebellar vermis and, additionally, in the pituitary gland. A distinct positive reaction was observed in all cortical brain regions and the neurons of the basal ganglia, the hypothalamus, the hippocampus and the cerebellar vermis but in the brainstem, relatively weak staining of neurons was seen. The reaction presented as a mainly diffuse cytoplasmic staining of the perikaryon of the neurons, and often axons and dendrites were also visualised. In addition, the immunoreactivity was present in the white matter. In the pituitary gland, however, distinct immunopositive cells were observed, with a prominent heterogeneity. The immunohistochemical findings were supported by the toxicological data. This immunostaining technique can be used as evidence of intake or even poisoning with MDMA and/or MDA and can be an interesting tool in forensic practice when the usual samples for toxicological analysis are not available. Furthermore, this method can be used to investigate the distribution of these substances in the human body.

Keywords:

3,4-Methylenedioxymethamphetamine (MDMA), 3,4-Methylenedioxyamphetamine (MDA), Immunohistochemistry, Human brain, Pituitary gland

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s004140100204

Issue: Volume 114, Number 6

Date: July 2001

Pages: 352 - 356

One fatal and seven non-fatal cases of 4-methylthioamphetamine (4-MTA) intoxication: clinico-pathological findings

E. A. De Letter ^A1, Vera A. E. Coopman ^A2, Jan A. C. M. Cordonnier ^A2, M. H. A. Piette ^A1

^A1 Department of Forensic Medicine, Ghent University, J. Kluyskensstraat 29, 9000 Ghent, Belgium e-mail: Michel.Piette@rug.ac.be, Tel.: +32-9-2649291, Fax: +32-9-2649495
^A2 Toxicological Laboratory, Chemiphar NV, L. Bauwensstraat 4, 8200 Bruges, Belgium

Abstract:

Abstract We present a case history involving one fatal and seven survived cases of intoxication with 4-methylthioamphetamine (4-MTA), also called para-methylthioamphetamine (p-MTA) or methylthioamphetamine (MTA), a relatively new amphetamine analogue. Two of the seven survivors required a 24-h-period of observation in hospital. This report proves once again that the new amphetamine designer drugs are not without danger, as is thought by many young people. In addition, individually different subjective reactions are described. Finally, the medico-legal implications of new, as yet unregistered drugs are discussed.

Keywords:

Keywords Drug abuse · 4-MTA · 4-Methylthioamphetamine · Amphetamines · Intoxication

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s004140050176

Issue: Volume 111, Number 6

Date: October 1998

Pages: 305 - 308

Identification of famprofazone ingestion

F. Musshoff ^A1, T. Kraemer ^A2

^A1 Institute of Legal Medicine, Friedrich-Wilhelms-University, Stiftsplatz 12, D-53111 Bonn, Germany
^A2 Institute of Pharmacology and Toxicology, Department of Toxicology, D-66421 Homburg/Saar, Germany

Abstract:

Abstract After a traffic accident a 32-year-old man was suspected of having previously taken an illegal drug. An immunochemical screening procedure revealed positive results for amphetamines in both urine and blood samples. The preliminary test was confirmed by GC/MS and both amphetamine and methamphetamine were found in both body fluids. However, the man denied any use of drugs but claimed to have taken four tablets of Gewodin. One of the ingredients, famprofazone, undergoes metabolic conversion to amphetamine and methamphetamine. Using GC/ MS the ingestion of famprofazone was verified by identification of the unchanged parent compound in the urine sample.

Keywords:

Key words Famprofazone · Amphetamine · Methamphetamine · Drug abuse · GC/MS analysis

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

Issue: Volume 113, Number 3

Date: May 2000

Pages: 168 - 170

Deaths associated with MBDB misuse

N. Carter ^A1, G. N. Rutty ^A1, C. M. Milroy ^A1, A. R. W. Forrest ^A2

^A1 Department of Forensic Pathology, University of Sheffield, The Medico-Legal Centre, Watery Street, Sheffield S37ES, UK
^A2 Department of Clinical Chemistry, Royal Hallamshire Hospital, Sheffield, UK e-mail: C.M.Milroy@Sheffield.ac.uk, Tel. +44-114-2738721, Fax +44-114-798942

Abstract:

Abstract The use of phenethylamines in the dance scene is now well established. Apart from amphetamine, the commonest phenethylamine encountered in clinical and forensic settings is 3,4-methylenedioxymethamphetamine (MDMA) commonly known as ecstasy. Other phenethylamines, which have similar effects are encountered, such as 3,4-methylenedioxyethylamphetamine (MDEA) and their use has resulted in death. We report two deaths associated with another less commonly encountered member of the group, N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine (MBDB), also known as Methyl-J and Eden.

Keywords:

Key words N-methyl-1-(3 · 4-methylenedioxyphenyl)-2-butanamine · MBDB Ecstasy · Eden Death

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International Journal of Legal Medicine

Publisher: Springer-Verlag Heidelberg

ISSN: 0937-9827

DOI: 10.1007/s00414-002-0290-2

Issue: Volume 117, Number 1

Date: February 2003

Pages: 2 - 9

Archives of Toxicology

Publisher: Springer-Verlag Heidelberg

ISSN: 0340-5761

Issue: Volume 68, Number 3

Date: March 1994

Pages: 210 - 211

Ethylmorphine concentrations in human samples in an overdose case

P. Kintz ^A1, C. Jamey ^A1, P. Mangin ^A1

^A1 Institut de Medecine Legale, 11 Rue Humann, 67085 Strasbourg Cedex, France FR

Abstract:

Capillary gas chromatography coupled to mass spectrometry was employed to quantify ethylmorphine in biological fluids and tissues in a death attributed to oral ethylmorphine ingestion. The femoral blood concentration of the drug was 488 ng/ml. Hair analysis revealed the individual's drug abuse pattern, particularly the switching from heroin to ethylmorphine.

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