Analysis of LSD in human body fluids and hair samples applying ImmunElute columns.

Immunoaffinity extraction units (LSD ImmunElute) are commercially available for the analysis of lysergic acid diethylamide (LSD) in urine. The ImmunElute resin contains immobilized monoclonal antibodies to LSD. We applied the ImmunElute procedure to serum and also to human hair samples. For hair analysis the samples were first extracted with methanol under sonication. The extracts were then purified using the ImmunElute resin. LSD analysis was carried out with HPLC and fluorescence detection. The immunoaffinity extraction provides highly purified extracts for chromatographic analysis. The limit of detection (signal-to-noise ratio = 3) has been determined to be < 50 pg regardless of which sa…

Development of a new multi-analyte assay for the simultaneous detection of opioids in serum and other body fluids using liquid chromatography-tandem mass spectrometry.

Abstract A liquid chromatography–tandem mass spectrometry method using electrospray ionization in positive ionization mode was developed for the simultaneous detection of multiple opioid-type drugs in plasma. The presented assay allows the quantitative determination of alfentanil, buprenorphine, codeine, desomorphine, dextromethorphan, dextrorphan, dihydrocodeine, dihydromorphine, ethylmorphine, fentanyl, hydrocodone, hydromorphone, methadone, morphine, naloxone, naltrexone, oxycodone, oxymorphone, pentazocine, pethidine, pholcodine, piritramide, remifentanil, sufentanil, and tramadol as well as the metabolites 6-monoacetylmorphine, bisnortilidine, morphine-3-glucuronide, morphine-6-glucuro…

Cumyl-PEGACLONE: A comparatively safe new synthetic cannabinoid receptor agonist entering the NPS market?

Detection of the synthetic drug 4-fluoroamphetamine (4-FA) in serum and urine.

Abstract 4-Fluoroamphetamine (4-FA) was detected in the blood and urine of two individuals suspected for driving under the influence (DUI). The test for amphetamines in urine subjected to immunoassay screening using the CEDIA DAU assay proved positive. Further investigations revealed a 4-FA cross-reactivity of about 6% in the CEDIA amphetamine assay. 4-FA was qualitatively detected in a general unknown screening for drugs using GC/MS in full scan mode. No other drugs or fluorinated phenethylamines were detected. A validated GC/MS method was established in SIM mode for serum analysis of 4-FA with a limit of detection (LOD) of 1 ng/mL and a lower limit of quantification (LLOQ) of 5 ng/mL. Int…

Identification of Potential Distinguishing Markers for the Use of Cannabis-Based Medicines or Street Cannabis in Serum Samples

Increasing prescription numbers of cannabis-based medicines raise the question of whether uptake of these medicines can be distinguished from recreational cannabis use. In this pilot study, serum cannabinoid profiles after use of cannabis-based medicines were investigated, in order to identify potential distinguishing markers. Serum samples after use of Sativex®, Dronabinol or medical cannabis were collected and analyzed for 18 different cannabinoids, using a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Analytes included delta-9-tetrahydrocannabinol, 11-hydroxy-tetrahydrocannabinol, 11-nor-9-carboxy-tetrahydrocannabinol, cannabidiol, cannabinol, cannabigerol, …

Determination of the enantiomeric composition of amphetamine, methamphetamine and 3,4-methylendioxy-N-methylamphetamine (MDMA) in seized street drug samples from southern Germany.

Amphetamine (speed), methamphetamine (crystal meth), and 3,4-methylenedioxy-N-methylamphetamine (MDMA, ecstasy) represent the most frequently abused amphetamine-type stimulants (ATS). Differences in pharmacological potency and metabolism have been shown for the enantiomers of all three stimulants. Legal consequences in cases of drug possession may also differ according to the German law depending on the enantiomeric composition of the seized drug. Therefore, enantioselective monitoring of seized specimens is crucial for legal and forensic casework. Various kinds of samples of amphetamine (n = 143), MDMA (n = 94), and methamphetamine (n = 528) that were seized in southern Germany in 2019 and…

Einfluss von THC auf die Glukoseaufnahme im Rattenhirn und das motorische Verhalten

Hautblasenbildung durch lokale Barbituratwirkung in der frühen Postmortalphase

Eine 31 Jahre alte Frau wurde tot mit anhangender Infusionsflasche und peripherem Venenzugang im Bereich des linken Handruckens aufgefunden. Bei der Leichenschau fielen ausgepragte Hautblasen mit seros-blutigem Inhalt sowie eine extreme Darstellung des Hautvenennetzes an der gesamten linken oberen Extremitat auf. Autolyseerscheinungen waren bei kurzer postmortaler Liegezeit (maximal ca. 24 h) nicht nachweisbar. Bei der toxikologischen Untersuchung wurde das bestimmungsgemas als Kurznarkotikum eingesetzte Barbiturat Thiopental nachgewiesen. Auffallig war eine hohe Thiopentalkonzentration in der Hautblasenflussigkeit von 1025 μg/ml, die die Konzentration im Oberschenkelvenenblut (5,35 μg/ml) …

Detection of  9-Tetrahydrocannabinol and Amphetamine-Type Stimulants in Oral Fluid Using the Rapid Stat  Point-of-Collection Drug-Testing Device

The Rapid Stat assay, a point-of-collection drug-testing device for detection of amphetamines, cannabinoids, cocaine, opiates, methadone, and benzodiazepines in oral fluid, was evaluated for cannabis and amphetamine-type stimulants. The Rapid Stat tests (n = 134) were applied by police officers in routine traffic checks. Oral fluid and blood samples were analyzed using gas chromatography-mass spectrometry (GC-MS) for Delta(9)-tetrahydrocannabinol, amphetamine, methamphetamine, methylenedioxymethamphetamine, methylenedioxyethylamphetamine, and methylenedioxyamphetamine. The comparison of GC-MS analysis of oral fluid with the Rapid Stat results for cannabis showed a sensitivity of 85%, a spec…

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…

False-positive LSD testing in urine samples from intensive care patients.

Unexpected positive results for lysergic acid diethylamide (LSD) were found in urine samples from 12 patients in an intensive care unit in a routine screening using the CEDIA DAU assay. None of these test results could be confirmed by high-performance liquid chromatography analysis, but all samples contained the mucolytic drug ambroxol. Further studies demonstrated that ambroxol exhibits a significant cross-reactivity in the CEDIA DAU LSD assay. Therefore, positive LSD results obtained with the CEDIA DAU assay have to be critically evaluated, particularly during the cold season, when infections of the respiratory tract often result in more frequent use of mucolytic medications.

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.

Concentrations of  9-Tetrahydrocannabinol and 11-Nor-9-Carboxytetrahydrocannabinol in Blood and Urine After Passive Exposure to Cannabis Smoke in a Coffee Shop

Cannabinoid concentrations in blood and urine after passive exposure to cannabis smoke under real-life conditions were investigated in this study. Eight healthy volunteers were exposed to cannabis smoke for 3 h in a well-attended coffee shop in Maastricht, Netherlands. An initial blood and urine sample was taken from each volunteer before exposure. Blood samples were taken 1.5, 3.5, 6, and 14 h after start of initial exposure, and urine samples were taken after 3.5, 6, 14, 36, 60, and 84 h. The samples were subjected to immunoassay screening for cannabinoids and analyzed using gas chromatography-mass spectrometry (GC-MS) for Delta(9)-tetrahydrocannabinol (THC), 11-nor-hydroxy-Delta(9)-tetra…

Einfluss von THC auf die Glukoseaufnahme und den zellulären Energiestatus im Rattenhirn und auf die Impulskontrolle

Extensive phytocannabinoid profiles of seized cannabis and cannabis-based medicines – Identification of potential distinguishing markers

As the frequency of cannabis-based therapy increases, the ability to distinguish intake of cannabis-based medicines from recreational cannabis use becomes desirable. Minor cannabinoids have been suggested to indicate recreational cannabis use in biological matrices but are unreliable when presumably also present in directly plantderived medicines. Thus, for therapeutics such as medical cannabis, Sativex® and Dronabinol, a more thorough investigation of cannabinoid profiles is required to identify possible distinguishing markers. In this study, 16 phytocannabinoids were quantified in samples of seized and medical cannabis, Sativex® and Dronabinol from two different manufacturers, using a val…

Zur forensischen Beweisführung bei einer Silikonembolie durch Kombination histologischer und toxikologischer Untersuchungsmethoden

Am Beispiel eines Falles aus dem Obduktionsgut des rechtsmedizinischen Institutes Frankfurt werden diagnostische Moglichkeiten zum Nachweis einer Silikonembolie aufgezeigt. Es handelt sich um den Fall einer 21 Jahre alten Frau, die nach kosmetischen Injektionen von flussigem Silikon verstarb. Die histologischen Befunde (lichtoptisch leere Hohlraume im Weichteilgewebe des Gesases mit umgebender Zellreaktion, leere dilatierte Blutgefase in den Lungen sowie leere Blutgefase in den Glomerula der Nieren) und die Resultate der toxikologischen Untersuchungen (NMR-spektroskopischer und massenspektrometrischer Nachweis von Silikon im Nieren- und Lungengewebe) erlaubten es, in ihrer Kombination eine …

False-Positive LSD Drug Screening Induced by a Mucolytic Medication

Since the early 1990s, the ingestion of lysergic acid diethylamide (LSD) as an inexpensive alternative to amphetamine derivatives has once again become widespread (1)(2). Consequently, screening of LSD has gained importance in clinical routine. The drug screening of a patient with a severe craniocerebral trauma showed a positive LSD screening by the homogeneous immunoassay CEDIA® DAU LSD (Boehringer Mannheim). In spite of the 3-h half-life of LSD in plasma (3), the drug screening remained positive for several days. These samples …

In vitro production of GHB in blood and serum samples under various storage conditions

Abstract The in vitro production of GHB was observed in freshly collected, untreated whole blood samples using glass BD-Vacutainers and polypropylene S-monovettes. GHB concentrations were determined daily over a period of one week and after 3, 6 and 9 weeks again. Furthermore, the GHB concentration in 40 untreated random whole blood samples stored at 4 °C for a longer period of time (10 samples 12 month, 10 samples 24 month and 20 samples 36 month) was also determined. For comparison, the in vitro production of GHB in freshly collected and prepared serum samples was observed. GHB serum concentrations were determined three times over a period of one week and once again after six weeks. Sampl…

Direct ELISA kits as a sensitive and selective screening method for abstinence control in urine.

In 2009 cutoff values of assessment criteria to testify abstinence control in order to estimate driving ability were standardized in Germany. The cutoff values are lower than required in existing guidelines like SAMHSA and there is critical discussion about detection of low concentrations by using immunoassay, especially concerning amphetamines in urine (50 ng/ml). In this study Direct ELISA kits were tested for their applicability to identify the absence of amphetamines, cannabinoids, opiates, cocaine, methadone and benzodiazepines in urine. Results were confirmed by LC/MS or GC/MS analyses. Sensitivity, specificity, predictive values (positive as well as negative) and overall misclassific…

Der Keto-Diabur-Test 5000 als schnelle Diagnosehilfe beim Tod durch Unterkühlung

Der Keto-Diabur Test 5000 (Boehringer Mannheim) ist ein Schnelltest zur Bestimmung von Glucose- und Ketonkorpern im Urin. Mit diesem routinemasig durchgefuhrten Test wurden bei der Obduktion von drei eindeutig infolge Erfrierens bzw. Unterkuhlung verstorbener Personen stark positive Resultate fur Ketone erhalten, ohne das gleichzeitig die Glucose im Harn erhoht war. Mittels Headspace-Gaschromatographie konnten in allen drei Urinproben uberhohte Aceton-Konzentrationen von 140, 1004 und 2703 mg/L festgestellt werden. Ein stark positiver Keto-Diabur-Test 5000 bei normalem Glucose-Befund im Urin kann somit als diagnostische Hilfe zur Erkennung eines Todes infolge Unterkuhlung dienen.

Simultaneous quantification of 18 different phytocannabinoids in serum using a highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method.

Abstract The potential therapeutic effects of various phytocannabinoids and the availability of multiple cannabis-based medicines make it desirable to have an analytical method that simultaneously quantifies a wide range of cannabinoids in blood, beyond delta-9-tetrahydrocannabinol and its metabolites. A liquid chromatography tandem mass spectrometry (LC-MS/MS) method for quantification of 18 phytocannabinoids and cannabinoid metabolites in serum was developed and validated. The method enables simultaneous detection of delta-9-tetrahydrocannabinol, cannabidiol, cannabinol, cannabigerol, cannabichromene, cannabicyclol, tetrahydrocannabivarin and cannabidivarin and their acidic precursors tet…

Einfluss chronischer THC-Gaben auf die zerebrale Glukoseaufnahme in der Ratte

Effects of tetrahydrocannabinol on glucose uptake in the rat brain

Δ9-Tetrahydrocannabinol (THC) is the psychoactive component of the plant Cannabis sativa and acts as a partial agonist at cannabinoid type 1 and type 2 receptors in the brain. The goal of this study was to assess the effect of THC on the cerebral glucose uptake in the rat brain. 21 male Sprague Dawley rats (12-13 w) were examined and received five different doses of THC ranging from 0.01 to 1 mg/kg. For data acquisition a Focus 120 small animal PET scanner was used and 24.1-28.0 MBq of [18F]-fluoro-2-deoxy-d-glucose were injected. The data were acquired for 70 min and arterial blood samples were collected throughout the scan. THC, THC-OH and THC-COOH were determined at 55 min p.i. Nine volu…

A fatal paramethoxymethamphetamine intoxication.

During the last years in Germany a marked increase in the use of amphetamines such as 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) has been observed. The use of these recreational drugs is especially common among young people participating in rave parties. Occasionally ring-methoxylated phenethylamine derivatives like paramethoxymethamphetamine (PMMA) or paramethoxyamphetamine (PMA) are found in street drugs offered as ecstasy. These compounds exhibit a higher toxicity than the methylenedioxyamphetamine derivatives. We report on the death of a 22-year-old man after the ingestion of ecstasy pills containing PMMA and PMA. The PMMA concentration in femoral blood was 0.85 mg/l. Besides PMA…