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Biomedical subjects

B Ludes

Publications and source records attributed to B Ludes.

At least 55 records · Page 3Linked to original sources

Detection of cannabis in oral fluid (saliva) and forehead wipes (sweat) from impaired drivers.

Saliva and sweat have been presented as two alternative matrices for the establishment of drug abuse. The noninvasive collection of a saliva or sweat sample, which is relatively easy to perform and can be achieved under close supervision, is one of the most important benefits in a driving-under-the-influence situation. Moreover, the presence of certain analytes in saliva is a better indication of recent use than when the drug is detected in urine, so there is a higher probability that the subject is experiencing pharmacological effects at the time of sampling. We developed an original procedure using gas chromatography-mass spectrometry to test for delta9-tetrahydrocannabinol (THC), the psychoactive ingredient of cannabis, in oral fluid and forehead wipes, collected with Sarstedt Salivettes and cosmetic pads, respectively. Blood, urine, oral fluid, and forehead wipes were simultaneously collected from 198 injured drivers admitted to an Emergency Hospital in Strasbourg, France. Of the 22 subjects positive for 11-nor-9-carboxy-THC (THCCOOH) in urine, 14 and 16 were positive for THC in oral fluid (1 to 103 ng/Salivette) and forehead wipe (4 to 152 ng/pad), respectively. 11-Hydroxy-THC and THCCOOH were not detected in these body fluids. Two main limitations of saliva and sweat are apparent: the amount of matrix collected is smaller when compared to urine, and the levels of drugs are higher in urine than in saliva and sweat. A current limitation in the use of these specimens for roadside testing is the absence of a suitable immunoassay that detects the parent compound in sufficiently low concentrations.

Adolescent↗

Doping control for beta-adrenergic compounds through hair analysis.

An original procedure was developed to simultaneously test beta2-agonists (salbutamol and clenbuterol) and beta-blockers (atenolol, acebutolol, pindolol, betaxolol, propranolol, timolol, sotalol, metoprolol, tertatolol, bisoprolol, labetalol and oxprenolol) in both human and animal hair. After decontamination with methylene chloride (2 times, 2 min), a 200 mg hair strand is pulverized in a ball mill. Then, a 100 mg portion is incubated overnight in 2 mL 0.1 N HCl, at 56 degrees C, in the presence of carteolol, which was used as an internal standard. After neutralization of the acid phase with 0.1 N NaOH, a 2 mL bicarbonate buffer (pH 8.6) is added to the preparation, which is then purified by solid-phase extraction with Isolute C18 columns. Drugs are derivatized using a mixture of trimethylboroxine-ethyl acetate for 15 min at 80 degrees C to form methaneboronate derivatives. Drugs are detected using GC/MS on an HP 6890-5973 system. A 4 microL portion of the derivatized extract is injected using a pulsed mode in a 30 m HP5 MS capillary column. Linearity was observed for all compounds in the range 25 pg/mg to 10 ng/mg. Limits of detection were in the range 2 to 10 pg/mg. At 1 ng/mg, recoveries were in the range from 37 to 100%, with a within-run precision of 5.9 to 14.1% (n = 8). The application of the method can be documented by the following examples: (1) Hair from asthmatic patients (n - 11), including two cases of asthma deaths, tested positive for salbutamol in the range of 27 to 210 pg/mg. (2) A 24-year-old swimmer who tested positive in urine for salbutamnol denied the results. Hair analysis confirmed salbutamol exposure, with a concentration of 71 pg/mg. (3) A shooting specialist was assumed to chronically use metoprolol (100 mg/daily during some periods). Hair concentration of metoprolol was 8.41 ng/mg. (4) An archery specialist was assumed to chronically use sotalol (80 mg/daily, during some periods). Hair concentration of sotalol was 261 pg/mg. (5) Hair from two calves revealed chronic exposure to clenbuterol, which was used to increase the mass of the animals at a concentration of 30 and 48 pg/mg.

Adrenergic beta-Agonists↗

Short Tandem Repeat loci analysis in forensic casework.

The development of polymerase chain reaction typing method has enhanced the possibilities of genetic identification of biological evidence samples for forensic caseworks. The authors report the use of multiplex amplification reactions associating several short tandem repeat loci. These amplification systems produce reliable results and a sufficient incrimination power to send the cases before the court.

Chromosome Mapping↗

Testing for alpha-chloralose by headspace-GC/MS. A case report.

A case is presented involving an acute fatality resulting from self-administered alpha-chloralose, a rodenticide. A 18-year-old man was found dead at home, with several stains of vomit on the carpet. An empty box of three bags of 5 g 100% alpha-chloralose (Corbeaux nuisibles, Rhône Poulenc) was found near the body. The compound was identified and quantified by headspace gas chromatography coupled to mass spectrometry. Alpha-chloralose was converted by concentrated sulphuric acid into chloral, a volatile compound, that was, after chromatography on a HP5-MS capillary column, identified by the following ions, m/z 82, 111 and 148. Peripheral blood concentration was 175.7 mg/l. Alpha-chloralose was quantified in several tissues, indicating kidney sequestration. No other drugs, including ethanol, were detected.

Adolescent↗

Testing for anabolic steroids in hair from two bodybuilders.

Two male bodybuilders were recently arrested by the French customs in Strasbourg (France) in possession of 2050 tablets and 251 ampoules of various anabolic steroids. It was claimed that the steroids were for personal use and not for trafficing as suggested by the police. Urine and hair specimens were collected from both suspects to clarify the claims. Nandrolone, stanozolol, testosterone and their corresponding metabolites were identified in the urine of both subjects. After decontamination, the hair was hydrolyzed by sodium hydroxide in presence of deuterated internal standards. After extraction with ethyl acetate and silylation, the drugs were identified by GC-MS in the electron impact mode. Hair from both males were positive for nandrolone (196 and 260 pg/mg), testosterone (46 and 71 pg/mg) and stanozolol (135 and 156 pg/mg), clearly indicating steroids abuse. Although not yet recognized by the International Olympic Committee, hair analysis may be a useful adjunct to conventional drug testing in urine from athletes.

Anabolic Agents↗

[Genetic imprints].

Explore the source record for details and available documents.

DNA Fingerprinting↗

Diatom analysis in victim's tissues as an indicator of the site of drowning.

The diagnosis of drowning is one of the most difficult in forensic pathology and previously we proposed criteria for a positive tissue analysis according to the qualitative and quantitative diatom investigations. In the positive cases, we studied the reliability of determining the site of drowning by comparing the diatom taxa found in the lung samples with those of the water samples or in the absence of these samples with the results of the water diatom monitoring programme set up in our region. In this study, we present two series of cases, the first is one of 20 corpses who died from accidental or suicidal drowning with known drowning site, and the second of 20 corpses for which the drowning site was unknown. The results showed that a concordance of the abundance of the diatom taxa in tissues compared to the site of drowning and their distribution relative to one another was 65% in the group where the site of drowning was known and 35% in the other group. The concordance of the individual distribution in the lungs of water diatom taxa may be an interesting method to guide the investigations for determining the site of drowning. The two limiting factors are the concentration of diatoms in the lungs and the development of a river monitoring programme in the district of the study.

Diatoms↗

Are cannabinoids detected in hair after washing with Cannabio shampoo?

Today, cannabis plants are used in shampoo preparations, in foodstuffs (e.g., oils, noodles, crackers, etc.), and in beverages (e.g., tea). These products often contain < 1% delta9-tetrahydrocannabinol (THC) in order to eliminate psychoactive effects, but some of them can include 1 to 3% of THC. Gas chromatography-mass spectrometry (GC-MS) analysis of Cannabio shampoo revealed the presence of THC (412 ng/mL) and two constituents of cannabis plants, cannabidiol (CBD, 4079 ng/mL) and cannabinol (CBN, 380 ng/mL). In order to verify if normal hygiene practices with Cannabio shampoo can result in positive tests for cannabinoids in hair, three subjects washed their hair with this shampoo once daily for two weeks. After this period, hair specimens were collected. In the three hair specimens, THC, CBD, and CBN were never detected within their limits of detection, 0.05, 0.02, and 0.01 ng/mg, respectively. We concluded that the use of Cannabio shampoo during normal hygiene practices cannot be considered as a source of potential contamination of hair. In a second experiment, drug-free hair specimens (200 mg) were incubated in 10 mL water/Cannabio shampoo (20:1, v/v) for 30 min, 2 h, and 5 h. After incubation, hair strands were washed with water and separated into two portions. One portion was extracted directly; the second was decontaminated with methylene chloride and then extracted. After an incubation period of 30 min, the analysis of hair by GC-MS did not reveal the presence of THC, CBD, and CBN in hair, regardless of whether the hair was decontaminated. After an incubation period of 2 h, specimens tested positive for CBD (0.11 ng/mg without decontamination and 0.10 ng/mg with decontamination) and CBN (0.02 ng/mg without decontamination and 0.02 ng/mg after decontamination). After an incubation period of 5 h, specimens tested positive for CBD (0.25 ng/mg without decontamination and 0.14 ng/mg after decontamination) and CBN (0.02 ng/mg without decontamination and 0.02 ng/mg after decontamination). In all cases, THC was never detected. Extensive but unrealistic use of Cannabio shampoo can cause drug-free hair to test positive for CBD and CBN but not for the primary psychoactive drug THC.

Animals↗

Identification of testosterone and testosterone esters in human hair.

In 1974, steroids were added to the list of doping agents banned by the International Olympic Committee because of their effects on the performance of the athletes. Testosterone and its esters promote the development of secondary male sexual characteristics and accelerate muscle growth. The mandatory test to detect testosterone abuse is to measure the ratio of testosterone to epitestosterone in the urine. However, because athletes can adjust their dosage to stay within the range permitted, there is a risk of test evasion. Therefore, we developed two original procedures to determine testosterone and its esters in human hair. First, testosterone was investigated in hair obtained from 26 control subjects. After decontamination with dichloromethane, 100 mg of hair was incubated in 1 M NaOH in the presence of 1 ng of testosterone-d3. After neutralization, the extract was purified using solid-phase extraction with Isolute C18 columns followed by liquid-liquid extraction with pentane. After silylation, testosterone was analyzed by gas chromatography-mass spectrometry. Concentrations were in the range 1.2 to 11.4 pg/mg with a mean value of 3.8 pg/mg. To distinguish exogenous abuse from endogenous levels, the incorporation of testosterone esters into hair was investigated. Preparation involved methanolic incubation to avoid the cleavage of the esters. In a panel of eight esters, it was possible to identify testosterone propionate, testosterone enanthate, and testosterone decanoate in the hair of two bodybuilders and one weight lifter. This new technology may find useful applications in anabolic abuse control.

Adolescent↗

Physiological concentrations of DHEA in human hair.

In 1974, steroids were added to the list of doping agents banned by the International Olympic Committee because of their effects on the performance of the athletes. Dehydroepiandrosterone (DHEA) is a steroid hormone naturally produced by the adrenal glands and by the ovaries. DHEA can be converted into other hormones, including estrogen and testosterone. In the United States, DHEA is classified as a nutritional supplement. This is not the case in France, where the drug is listed as a doping agent. As athletes can abuse DHEA to benefit from its conversion to testosterone, there is a need to establish the physiological range of DHEA concentrations in human hair. DHEA was investigated in hair obtained from 27 control subjects, including 15 males and 12 females aged 17-42 years. After decontamination with dichloromethane, 100 mg of hair was incubated in 1 M NaOH in presence of 1 ng of testosterone-d3. After neutralization, the extract was purified using solid-phase extraction with Isolute C18 columns and subsequent liquid-liquid extraction with pentane. After silylation, DHEA was analyzed by gas chromatography-mass spectrometry. Results were linear in the range 1-20 pg/mg. Relative extraction recovery was 91.6% with a limit of detection of 0.5 pg/mg. Concentrations were in the range 1.2-6.7 pg/mg (mean value of 4.3 pg/mg) and 0.5 to 10.6 pg/mg (mean value of 5.3 pg/mg) for the males and females, respectively. Extensive chromatographic procedures (two purification steps by solid-phase and liquid-liquid extraction, combined with injection of 4 microL through the column in pulsed mode) were analytical prerequisites for successful identification of DHEA in hair because of the low target concentrations. This new technology may find useful applications in anabolic abuse control.

Adjuvants, Immunologic↗

[Evidence of pesticide exposure by hair analysis].

The current report summarizes the development of an analytical method for the identification and the quantification of pesticides in hair by gas chromatography/mass spectrometry and its application to 75 real samples. Hair strands [table: see text] were obtained from wine workers exposed to one or more pesticides. After decontamination, hair were cut into small pieces and incubated overnight at 45 degrees C in methanol. The solvent was evaporated to dryness, the dry extract was redissolved in methanol and injected in a gas chromatography/mass spectrometry system. The detector was operated in electronic impact and in negative chemical ionization mode of detection (reactant gas: methane). In the first series of 75 hair specimens, obtained before the period of pesticide use, none of the 15 target compounds was detected. In the second series of 75 specimens, obtained from the same subjects but after the use of pesticides, 14 tested positive for 9 different pesticides.

Agrochemicals↗

[Norandrostenolone and noretiocholanolone: metabolite markers].

In order to evaluate the concentrations of norandrosterone (NA) and noretiocholanolone (NE) in human urine, nandrolone sulfate, nandrolone undecanoate, norandrostendiol and norandrostendione were administered, each to a different subject. The derivatized steroids were identified after pulsed mode injection by gas chromatography-mass spectrometry in electron impact mode, with selected ion monitoring. In the first case, NA and NE were detected in urine until 48 hours after oral administration of 22 mg of nandrolone sulfate. In the second case, urine specimens gave a positive response for NA and NE up to 8 months after intramuscular injection of 50 mg of nandrolone undecanoate. Finally, NA and NE were detected for 8 days after oral administration of 50 mg and 100 mg of norandrostendiol and norandrostendione. However, in contrast with nandrolone undecanoate, the ratio NA to NE was different according to the time of urine collection, after nandrolone, norandrostendiol and norandrostendione administration. From about 60 urinary samples obtained from both standard population and athletes, it was possible to characterize a physiological excretion of nandrolone metabolites. All concentrations were clearly under the proposed positive cut-off level (2 ng/ml) and the concentration of NA was always higher than the concentration of NE.

Administration, Oral↗

[Acute poisoning during substitution therapy based on high-dosage buprenorphine. 29 clinical cases--20 fatal cases].

OBJECTIVES: Buprenorphine has been an important advance in care for drug abusers, but the toxic risk may be fatal. We report here two original series of buprenorphine poisoning in opiate abusers on substitution therapy. PATIENTS: The first series included 20 males and 9 females, aged 20-35 years (mean = 27.5) with non-fatal poisoning. The second series included 20 subjects (19 males, 1 female) aged 14-48 years (mean = 26.6) with a fatal outcome. All subjects were opiate addicts taking high-dosage sublingual buprenorphine formulation as substitution therapy. RESULTS: Blood concentrations of buprenorphine were found in all cases to remain at a low level (1.0-2.3 ng/ml, m = 1.4 ng/ml, and 1.1-29.0 ng/ml, m = 8.4 ng/ml in non-fatal and fatal cases respectively). Almost all cases involved concomitant intake of psychotropic medications, especially benzodiazepines (18 non-fatal and 17 fatal cases). DISCUSSION: These observations confirm previously reported data on the danger of buprenorphine-benzodiazepine combinations. Intravenous injection of crushed tablets also appears to be a risk factor (8 deaths and 10 non-fatal poisonings). This series highlights the need for improvement in the recently developed French program for substitution therapy with high-dosage buprenorphine in heroin addicts.

Administration, Sublingual↗

Nicotine monitoring in sweat with a sweat patch.

In recent years, remarkable advances in sensitive analytical techniques have enabled the analysis of drugs in unconventional samples, such as sweat. In a study conducted with cigarettes smokers and nonsmokers, PharmChek sweat patches were applied to 29 subjects for 72 h. Nicotine was extracted in 5 ml methanol in the presence of 200 ng nicotine-d4, used as internal standard. After 20 min agitation, the methanolic solution was evaporated to dryness in the presence of 10 microl octanol to ensure nonvolatility of nicotine. Nicotine was determined using gas chromatography coupled to mass spectrometry after separation on a 30-m capillary HP5 MS column. The assay was linear in the range 50-2500 ng/patch, with an extraction recovery of 76+/-5%. Limit of detection was 10 ng/patch. Nicotine concentrations in sweat were not detected for the nonexposed nonsmokers (n = 8), 87 to 266 ng/patch for the passive smokers (n = 6) and 150 to 2498 ng/patch for the smokers (n = 15). This study demonstrated a useful application of the sweat patch for monitoring tobacco exposure.

Adult↗

Confirmation of oleander poisoning by HPLC/MS.

A non-fatal case of Nerium oleander (common oleander) self-poisoning in a 45-year-old female is presented. Initial symptoms were nausea and vomiting, abdominal pain, phosphenes, cardiovascular shock and sinus brady-cardia. Blood and urine were assayed for oleandrin, the major cardiac glycoside of N. oleander, using a highly specific HPLC/MS procedure. The blood concentration of oleandrin at admission was 1.1 ng/ml. This is the first report of an oleander intoxication ascertained by the mass spectrometric identification of oleandrin in blood. HPLC/MS appears to be the method of choice for the forensic-toxicological investigation of poisonings by cardiac glycosides.

Cardenolides↗

Codeine testing in sweat and saliva with the Drugwipe.

With the growing interest in drug testing within different sectors of society, there has become a need for drug assays that can be performed immediately at the site of specimen collection. Recently, Securetec (Ottobrunn, Germany) has introduced the Drugwipe, a non instrument-based, on-site immunodiagnostic assay for the detection of drugs on surfaces. Different tests are available for opiates, cocaine and cannabis. To document the applications of the Drugwipe "opiate" on human biological fluids, 60 mg codeine phosphate were orally administered to 6 subjects. First, sweat testing with the Drugwipe was studied. The wiping section of the kit was used to swab the forehead of the subjects for 10 s, at 1, 4, 9 and 24 h after codeine administration. At the same time, for each period, a sweat patch (Pharmchek, USA) was applied to the outer portion of the upper arm. Codeine was then quantified in the patch by GC/MS and the measured concentrations used as reference. In all subjects except one the Drugwipe tested positive for opiates, however with few false negative results. In the second part of the study, results of the Drugwipe were compared with those obtained by GC/MS for saliva. The tongue of the subjects was carefully wiped over a period 24 h, and at the same time a specimen of saliva collected. Although codeine could be detected using the Drugwipe, numerous false negative results were observed. Codeine tested positive by GC/MS but remained negative using the Drugwipe in several cases. This can be explained by a codeine concentration which was too low to show positive with the Drugwipe, interfering substances may be present in saliva or the sampling procedure is inadequate.

Adult↗