Postmortem distribution of dihydrocodeine and metabolites in a fatal case of dihydrocodeine intoxication.

A report of a fatal dihydrocodeine ingestion under substitution therapy is given. Quantitation of dihydrocodeine, dihydromorphine, N-nordihydrocodeine, dihydrocodeine-6-, dihydromorphine-6- and dihydromorphine-3-glucuronide was performed simultaneously after solid-phase extraction prior to HPLC analysis, and the analytes were detected using their native fluorescence. Postmortem concentrations of blood samples from different sampling sites as well as from liver, kidney and cerebrum are reported. A hair sample was investigated to prove long-term use of the substitute drug. Site-to-site differences of the analytes from blood samples were very small. The partition behavior of the opioid glucuro…

Report on Intrauterine Drug Exposure During Second Trimester of Pregnancy in a Heroin-Associated Death

A 17-year-old girl was found dead in a public toilet with fresh needle puncture marks. She was 18-20 weeks pregnant with a male fetus. Drug screening of her blood and urine indicated recent heroin use. Chronic drug use was confirmed by hair analysis. Amniotic fluid as well as fetal and maternal tissues and body fluids were analyzed by GC/MS and HPLC. All the fetal specimens were investigated, and the following levels of drugs were found: 6-monoacetyl-morphine (blood: 152 ng/g; amniotic fluid: 128 ng/g; brain: 140 ng/g; lung: 110 ng/g; liver: 2 ng/g; kidney: 40 ng/g), morphine (blood: 1360 ng/g; amniotic fluid: 604 ng/g; brain: 710 ng/g; lung: 1030 ng/g; liver: 2060 ng/g; kidney: 1100 ng/g),…

In vitro contamination of hair by marijuana smoke.

Abstract Background: The deposition of cannabinoids on/into hair from environmental smoke can be considered as a potential source of drug findings in hair. We studied external uptake of cannabinoids from marijuana smoke, investigating possible influencing factors on drug uptake and the efficiency of decontamination procedures. Methods: Strands of a natural hair sample were moistened with water, greased with sebum or sebum/sweat, or bleached or permed. Treated and untreated samples were exposed to marijuana smoke for 60 min. Aliquots of each hair strand were either kept unwashed or were washed with methanol, dichloromethane, or 5 g/L dodecyl sulfate in water. Cannabinoid concentrations in un…

Zum Suchtmittelnachweis in Haaren IV. Einfluß der Pigmentierung auf den Ofloxacingehalt in Haaren bei Meerschweinchen

Meerschweinchen (n = 6) mit dreifarbigen Fellarealen (Tricolor) wurde 3 Wochen lang Ofloxacin (1 mg/mL) im Trinkwasser verabreicht. Vor Versuchsbeginn und danach wurden Haarproben von schwarzen, rotbraunen und weisen Fellarealen jedes Tieres getrennt gesammelt und der Ofloxacin-Gehalt mittels HPLC bestimmt. Sowohl in nicht pigmentierten als auch in pigmentierten Haaren konnte die Wirksubstanz nachgewiesen werden. Die hochsten Gehalte lagen in schwarzen, die niedrigsten Konzentrationen in weisen Haarproben vor. Rotbraune Fellareale enthielten weniger Ofloxacin als schwarze Fellareale des gleichen Tieres. Bei niedrigen Dosierungen waren die Unterschiede des Ofloxacingehalts verschiedenfarbige…

Influence of Pigmentation on the Codeine Content of Hair Fibers in Guinea Pigs

Tortoise shell guinea pigs (n = 7) were administered codeine (1 mg/mL codeine-base) in their drinking water for 3 weeks. Black, reddish-brown and white hair was collected separately from each animal before and after treatment. The hair samples were analyzed by GC/MS. The experiment showed positive results for all hair fibers with large individual variability of drug incorporation. Low drug intake resulted in small differences of the drug content in hair fibers different in color, whereas in cases of high drug intake a strong influence of hair pigmentation on the analytical results was observed. The highest drug content was always found in black hair samples, non-pigmented hair showed the lo…

Preliminary approach to elucidate the role of pigment as a binding site for drugs and chemicals in anagen hair: differential uptake of 3 H-haloperidol by pigment-producing compared to non-pigment-producing cell lines

A striking difference was observed for cellular-bound drug in HaCaT and Sk-Mel-1 cells for a fixed drug exposure time of 72 h and varying 3H-haloperidol concentrations in the culture media. Drug uptake was dependent on drug concentration and linearly correlated for both the non-pigment- and the pigment-producing cells which however was different in magnitude. In an additional investigation the time course of drug uptake during 3H-haloperidol exposure (400 pmol/ml; 28 days) revealed increasing drug concentrations in the Sk-Mel-1 population, whereas drug concentrations in the keratinocytes reached a plateau within a short time period. In contrast to the HaCaT cells no tendency to saturation w…

Freie und glucuronidierte Cannabinoide im Urin - Untersuchungen zur Einschätzung des Konsumverhaltens

Spontanurinproben (n = 135) von ¶49 Cannabiskonsumenten wurden auf freie und glucuronidierte Tetrahydrocannabinolcarbonsaure (THCCOOH) mittels LC/MS/MS sowie auf freies und glucuronidiertes Tetrahydrocannabinol (THC) und 11-Hydroxytetrahydrocannabinol (11-OH-THC) mittels GC/MS direkt sowie nach enzymatischer Hydrolyse bis zu 10 Tagen nach dem letztmaligen Konsum untersucht. Die Einteilung des Konsumverhaltens erfolgte in schwer, moderat und leicht und orientierte sich an den Zuordnungskriterien neuerer, einschlagiger Publikationen. Die Konzentrationsangaben fur freie und glucuronidierte THCCOOH wurden auf 100 mg Kreatinin/dL Urin bezogen. In den Konsumentengruppen schwer, moderat und leicht…

Biochemical approach on the conservation of drug molecules during hair fiber formation

A biochemical concept for the endogenous incorporation of drug molecules into growing hair is presented. It is based on the principles of transport across biomembranes, on the principles of biotransformation and drug melanin affinity. The approach gives explanations for current observations in hair analysis, which up to date have not been understood sufficiently. Phenomena such as the ratio of parent drug to metabolite in hair, the dependence of incorporation on the physico-chemical properties of the drug, the independence of drug incorporation on active melanogenesis (incorporation into non-pigmented hair) as well as the dependence of drug content on hair pigmentation are elucidated.

Zum Suchtmittelnachweis in Haaren V. Auswirkung von Sonne, Regen und Wind auf den Drogengehalt in Kopfhaaren von Drogenkonsumenten – ein Pilotprojekt

Der Einflus von Sonne, Regen und Wind fuhrt zu Schadigungen der Haarfaser. Zur Prufung witterungsbedingter Einflusse auf den Drogengehalt wurden authentische Haarproben wahrend 3 Sommermonaten der Globalstrahlung bzw. der naturlichen Witterung ausgesetzt. Die Untersuchungen zeigten, das insbesondere die Konzentrationen an Morphin and 6-Acetylmorphin nach Exposition abgenommen hatten, wahrend die Gehalte an Cocain und Benzoylecgonin weniger beeinflust worden waren. Die Konzentration an Dihydrocodein nahm insbesondere nach Lichtalterung der Haarproben zu. Es wird vermutet, das neben Auswascheffekten, hydrolytischem und oxidativem Abbau der Substanzen die Schadigung des Haarpigments durch UV-L…

An investigation of the stability of free and glucuronidated 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid in authentic urine samples.

Preanalytical stability of a drug and its major metabolites is an important consideration in pharmacokinetic studies or whenever the analyte pattern is used to estimate drug habits. Firstly, the stability of free and glucuronidated 11-nor-delta9-tetrahydrocannabinol-9-carboxylic acid (THCCOOH, THCCOOglu) in authentic urine samples was investigated. Random urine samples of cannabis users (n = 38) were stored at -20, 4, and 20 degrees C up to 15 days and up to 5 days at 40 degrees C, and alterations of the analyte pattern during storage were followed by liquid chromatography-tandem mass spectrometry. Secondly, the influence of pH (range 5.0-8.0) on the stability of the analytes was studied us…

Zum Suchtmittelnachweis in Haaren

Es wird ein Konzept zum Einbau organischer Substanzen in das wachsende Haar vorgestellt. Es basiert auf den Regeln des Stofftransportes durch Zellmembranen, den Prinzipien des Fremdstoffmetabolismus und der bekannten Melaninaffinitat vieler Substanzen. Die interdisziplinare Betrachtung und die Anwendung biochemischer Mechanismen auf die Vorgange und Zellpopulationen im Haarfollikel kann die Konservierung der organischen Substanzen infolge der Keratinisierungsprozesse und einige bisher noch nicht vollig verstandene Phanomene, wie das Konzentrationsverhaltnis von Muttersubstanz zu Metaboliten im Haar, erklaren. Nachvollziehbar wird auch, das der Einbau von Substanzen einerseits unabhangig von…

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

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-m…

Influence of sample preparation on analytical results: drug analysis [GC/MS] on hair snippets versus hair powder using various extraction methods

The comparison of aqueous extraction methods and hair extraction by organic solvents performed on hair powder as well as on hair snippets of the same sample revealed different qualities of the procedures. Qualitative and quantitative results by the same derivatization step and GC/MS detection demonstrated, that the risk of missing a drug substance is higher using hair snippets than after drug extraction on pulverised hair. Drug recovery for opiates, cocaine and benzoylecgonine from hair was found to be best in aqueous solvents or in methanol extracts. The results are discussed under the aspects of solid-phase extraction, the hair sample representing an inhomogenous material. The localisatio…

Zur präanalytischen Phase chemisch-toxikologischer Untersuchungen

Urin, ein bevorzugtes Untersuchungsmaterial fur ein Drogenscreening, ist haufiger als andere biologische Proben Manipulationen unterworfen. Da sich in jungster Zeit ein regelrechter Wettstreit zwischen Suchtmittelkonsument und Analytik entwickelt hat, wurden die wichtigsten Informationen zu dieser Thematik zusammengestellt. Als bedeutendste in-vivo Manipulation wird die Aufnahme groser Flussigkeitsmengen erachtet, und zur Erkennung einer Verdunnung des Urins wird die Bestimmung des Kreatininwertes empfohlen. Die Aussagekraft dieses Parameters an Spontanurinproben wird diskutiert, und Alternativen beim Vorliegen niedriger Kreatininwerte werden diskutiert.

Perspiration versus saliva--basic aspects concerning their use in roadside drug testing.

Various aspects concerning the practical application and forensic interpretation of data obtained by saliva drug testing and drug monitoring from the skin surface are discussed. Basic information on the composition of saliva and skin secretions and their particular transport mechanisms, as far as known, are given. For drugs of abuse secretion into saliva is suggested to be by passive diffusion and to depend on lipid solubility, pKa, plasma protein binding and on the pH of saliva. Drug molecules from blood are considered to reach the skin surface by various routes such as by sweat and sebum as well as by inter- and/or transcellular diffusion. The role of the stratum corneum as a temporary dr…

A preliminary study on the stability of benzodiazepines in blood and plasma stored at 4 degrees C.

An approach to determine the stability of benzodiazepines and some of their metabolites (n = 13) by means of a routinely applied gas chromatographic method using electron capture detection was made in this preliminary study. Validation data of the method are given. Spiked blood and plasma samples were stored at 4 degrees C and analysed at selected times up to 240 days. The concentrations of all analytes had decreased to at least 60% of the original levels at the end of the observation period. A clear pattern of breakdown could not be established. The data obtained suggest that results from long-term stored samples should be interpreted cautiously. Further investigations concerning the stabi…

Preliminary Practical Findings on Drug Monitoring by a Transcutaneous Collection Device

A noninvasive and nonocclusive skin patch (Sudormed™) was investigated for the systematic collection of drugs of abuse over a period of several days. First, the applicability and user friendliness were tested by volunteers. The permeability of the polyurethane dressing from the outside to the inside for an aqueous solution was shown by incubating the outside layer with Rhodamine B. No fluorescence could be detected in the cotton pad beneath. A single dose experiment using theophylline as a model compound showed that there was a delay in time before the substance could be determined in the pad. The drug content decreased with increasing time of patch application. When eight volunteers partic…

Partition coefficient, blood to plasma ratio, protein binding and short-term stability of 11-nor-Delta(9)-carboxy tetrahydrocannabinol glucuronide.

11-Nor-Delta(9)-carboxy tetrahydrocannabinol glucuronide (THCCOOglu) is a major metabolite of tetrahydrocannabinol in blood. Despite its mass spectrometric identification already in 1980, further physicochemical data of THCCOOglu have not been established. Therefore, the octanol/buffer partition coefficient P and the blood to plasma ratio b/p for THCCOOglu concentrations of 100 and 500ng/ml were investigated. Protein binding of the glucuronide was established from spiked albumin solutions at a level of 250ng/ml as well as from authentic samples. The data were compared to those of 11-nor-Delta(9)-carboxy tetrahydrocannabinol (THCCOOH). In addition, the short-term stability of THCCOOglu in pl…

Preliminary approach to elucidate the role of pigment as a binding site for drugs and chemicals in anagen hairs: pigments as carriers for 3 H-haloperidol in HaCaT/Sk-Mel-1 co-cultures

In view of the melanin-binding characteristics of haloperidol and its differential uptake by pigment- and non-pigment-producing cells, a co-culture of HaCaT with Sk-Mel-1 cell lines was performed to investigate whether melanosomes act as carriers for drug molecules associated with the pigments. Initially, HaCaT and Sk-Mel-1 cells were separately cultivated in the presence of 3H-haloperidol (400 pmol/ml medium ) for 28 days followed by subsequent co-cultivation in the absence of 3H-haloperidol for 5 days. The transfer of pigments into the keratinocytes during co-culture was confirmed by transmission electron microscopy. After the co-culture experiments a striking increase (or = 50%) of 3H-ha…

An In Vitro Experiment for Postmortem Vascular Permeation. The Passage of Morphine and Morphine Glucuronides Across a Vascular Wall

A venous blood sample taken at autopsy cannot be considered to represent the antemortem blood concentration of a particular substance. Autolytic processes cause disintegration and increasing permeability of the physiological and anatomical barriers such as vascular walls and lead to changes in substance concentrations. In the present study, the experimental design represents an in vitro postmortem simulation of a drug substance crossing a venous wall. The postmortem behavior of morphine, morphine-3- and morphine-6-glucuronide was investigated. A Chien-Valia-diffusion chamber with a patch of inferior vena cava as diffusion barrier was used. For optimal simulation of postmortem events, vein s…

On cosmetically treated hair — aspects and pitfalls of interpretation

Popular hair cosmetic treatments like bleaching or permanent waving were found to affect the stability of incorporated drugs and to cause alterations of the fibers at an ultrastructural level. This may result in a partial or complete loss of drug substances, depending on the particular drug molecule and on its concentration prior to the cosmetic treatment. Moreover, from literature, there is some evidence that drug molecules are not only incorporated into the growing fiber by passive diffusion from blood into the matrix cells and melanocytes, but that the substances enter the hair also via perspiration such as sweat and sebum. Since permed and bleached hair shows an enhanced sorption capaci…

Short-Term Stability of Lysergic Acid Diethylamide (LSD), N-Desmethyl-LSD, and 2-Oxo-3-hydroxy-LSD in Urine, Assessed by Liquid Chromatography–Tandem Mass Spectrometry

Lysergic acid diethylamide (LSD) is one of the most potent hallucinogenic agents known. Recently, data on emergency department episodes related to the use of drugs commonly thought as “club drugs” have also included LSD (1). Confirmation of LSD use by testing biological fluids is still an analytical challenge because of its extensive, rapid metabolism and its instability (2)(3)(4). After ingestion of a typical street dose (40–120 μg), the concentration of LSD in urine falls to <1 μg/L within a few hours (2)(5)(6). Recently, N -desmethyl-LSD (nor-LSD) and 2-oxo-3-hydroxy-LSD (O-H-LSD) have been identified as LSD metabolites in human urine (7)(8). Measured nor-LSD concentrations were reported…

Stability of Cannabinoids in Hair Samples Exposed to Sunlight

It has been well recognized that hashish and marihuana lose potency during storage because of a decrease in the content of tetrahydrocannabinol (THC), which is the major psychoactive constituent of cannabis (1). The effect of oxygen on stored plant and resin materials or solutions of pure cannabinoids seems much less significant than that of higher temperatures (≥37 °C) or light (2)(3)(4)(5)(6)(7). A few data are available on the stability of THC and major metabolites in blood (8)(9)(10)(11). However, the stability of cannabinoids in the hair shaft has not been addressed, although scalp hair represents one of the most exposed parts of the body. Therefore, a study was performed to elucidate …

Plasma-to-Blood Ratios of Congener Analytes

A preliminary study on the distribution of morphine and its glucuronides in the subcompartments of blood.

[Abstract ] The distribution of morphine, morphine-3-glucuronide (M3G), and morphine-6-glucuronide (M6G) in whole blood, plasma, and packed erythrocytes was studied. Parameters investigated were the hematocrit values (10, 42, 44, and 71%) and the water content of the samples. The blood-to-plasma ratio of morphine concentrations was unaffected by variations in hematocrit and water content, whereas the corresponding ratios for M3G and M6G were strongly influenced. Ratios were 0.53 to 0.65 and 0.52 to 0.62 in specimens with average hematocrit values (42 and 44%, respectively), and the ratios were 0.81 or 0.89 (hematocrit 10%) and 0.27 or 0.28 (hemalocrit 71%) in blood samples with different he…

Diskurs zu Morphinbefunden im Autopsieblut nach palliativer Schmerzbehandlung

Nach einer Schmerzbehandlung stellt die Beurteilung von Morphin- und seinen Glucuronidkonzentrationen am Leichenblut hohe Anforderungen an einen forensischen Gutachter. Im vorliegenden Beitrag werden wichtige physikochemische Eigenschaften sowie pharmakokinetische Daten der Analyte zusammengestellt und eine kurze Ubersicht des Wissenstands uber die Pharmakokinetik von Morphin gegeben. Die Durchsicht klinischer Untersuchungen ergab eine grose Bandbreite therapeutischer Morphindosierungen und eine hohe Variabilitat aller pharmakokinetischer Parameter. Auf die Auswirkungen einiger Einflussfaktoren wie Applikationsweg, Applikationsfrequenz, ¶Alter und Allgemeinzustand sowie Leber- und Nierenfun…

Ultrastructural alterations and environmental exposure influence the opiate concentrations in hair of drug addicts

Hair samples were taken at autopsy from the head of 1 male and 1 female subject both known as drug abusers. Some of the strands were bleached by in-vitro cosmetic treatment. The bleached hair as well as the original hair samples were partly exposed to water or soil prior to further investigations and drug monitoring. The exposure times were 4 weeks or 6 months for water and 6 months for soil. The hair fibers were examined by transmission electron microscope (TEM) and by scanning electron microscope (SEM) investigations. The electron microscope studies confirmed that all experimental conditions had produced morphological alterations in the hair fibers. After exposure to water or to soil for …

Stability of Morphine, Morphine-3-Glucuronide, and Morphine-6-Glucuronide in Fresh Blood and Plasma and Postmortem Blood Samples

The present study was designed to determine the stability of morphine and its glucuronides in spiked fresh blood and plasma from live individuals as well as in four authentic postmortem blood specimens for a time interval of up to six months. The samples were stored in glass vials at -20 degrees C, 4 degrees C, and 20 degrees C. Additionally, spiked samples were exposed to light through window glass and subjected to a forced-degradation study at 40 degrees C. Data were established using solid-phase extraction and high-performance liquid chromatography coupled to atmospheric pressure ionization mass spectrometry for isolation and quantitation, providing a sensitive and specific detection met…

Saliva testing after single and chronic administration of dihydrocodeine.

In the present study, concentrations of dihydrocodeine and its metabolites in saliva and serum were compared after single low-dose and chronic high-dosage administration of the drug. In the first investigation, blood and saliva were collected periodically from six subjects after oral administration of 60 mg dihydrocodeine. In the second study, 20 subjects on oral dihydrocodeine maintenance provided single samples of blood and saliva simultaneously. Serum protein binding of salivary analytes and their recovery from the adsorbing material of the collection device as well as pH values of saliva samples were determined. The fluids were analyzed for dihydrocodeine and the major metabolites by hi…

Zum Suchtmittelnachweis in Haaren. VII. Untersuchungen bei Methadonpatienten unter konstanter Wirkstoffeinnahme

Bei 18 Patienten unter Methadonsubstitutionstherapie mit konstanter Dosis konnte bei Berucksichtigung des individuellen Haarwachstums keine zufriedenstellende Korrelation (r = 0,602) zwischen Aufnahmemenge und Methadongehalt im Haar gefunden werden. Die uber 4 Tage durchgefuhrten Sammeltests mit Perspirationspflaster bestatigten das Vorhandensein von Methadon und seinem Hauptmetaboliten auf der Hautoberflache und zeigten bei 6 Personen einen Beikonsum, dessen Substanzspektrum teilweise auch in den untersuchten Haarabschnitten nachzuweisen war.

Experimental Investigations on Hair Fibers as Diffusion Bridges and Opiates as Solutes in Solution

Diffusion experiments were performed using clipped hair fibers as diffusion bridges and aqueous solutions of morphine, codeine and dihydrocodeine. Natural as well as predamaged hair fibers were investigated. The test series were conducted at ambient temperature and at high humidity. After 312 or 372 hours the middle segments of the strands were clipped, washed and analyzed by GC/MS. Only when virgin hair samples were used the solutes passed along the fiber at full length resulting in a positive immunological finding at the end of the diffusion bridge. Most of the washing fluids were positive for opiates. All centerpieces had a high opiate content. The opiate concentration in damaged hair wa…

Stability of 11-Nor-Δ9-carboxy-tetrahydrocannabinol Glucuronide in Plasma and Urine Assessed by Liquid Chromatography-Tandem Mass Spectrometry

Abstract Background: Unconjugated 11-nor-Δ9-carboxy-tetrahydrocannabinol (THCCOOH) in blood and urine has been proposed as a valuable marker, but the glucuronide (THCCOOglu) is present in considerably higher concentrations than the parent drug. Acyl glucuronides have been shown to be potentially reactive conjugates, which may affect the in vitro metabolite pattern. Methods: Extraction procedures and a liquid chromatography-tandem mass spectrometry assay were developed and validated to investigate the stability of THCCOOglu in urine and plasma. Plasma and urine samples with added THCCOOglu were stored at −20, 4, 20, and 40 °C up to 10 days. Results: The glucuronide was stable at −20 °C in bo…