Determination of cocaine in human hair by gas chromatography/mass spectrometry

Summary A qualitative method for the determination of cocaine alone without its metabolites in human hair by gas chromatography/mass spectrometry (GC/MS) was developed. The assay used helium as carrier gas, a 30-m bonded phase fused silica OV-1 capillary column, and solid injection at 290°C evaporator temperature.The cocaine concentrations in hair were determined also by radio-immunoassay (RIA). The values obtained are the sum of cocaine and its metabolites.Both GC/MS and RIA meet the requirements for the determination of drug abuse by two different methods in forensic science.     

Determination of methadone in human hair by radioimmunoassay

Summary The concentrations of methadone in human hair were measured. The washed hair was cut in 1-mm pieces approximately, then incubated overnight at 45°C with 0.1 m HCl. The extracts were alcalized by 1 m NaOH and diluted by phosphate buffer, pH 7.4. The methadone concentrations were determined by radioimmunoassay. The method is simple, rapid, and practicable for routine determination.     

Methadone concentrations in human hair of the head,axillary and pubic hair

Summary The concentrations of methadone and its metabolites in the hair of the head, axillary and pubic hair obtained from patients receiving a daily maintenance doses, were determined. Comparison of the concentrations provides the highest values in the axillary hair, followed by pubic hair and the hair of the head.     

Concentrations of Δ9-tetrahydrocannabinol,cocaine and 6-monoacetylmorphine in hair of drug abusers

Hair samples taken from 850 individuals with presumed drug abuse were tested simultaneously forΔ⁹tetrahydrocannabinol (THC), cocaine, heroin, the primary heroin metabolite 6-monoacetylmorphine (6-MAM) and morphine. The drugs were extracted with methanol under sonication. Compared to other extraction procedures this solvent extraction technique provides high extraction yields and less experimental effort. The analyses were carried out using gas chromatography - mass spectrometry (GCMS) in selected ion monitoring (SIM) mode. This procedure allows the simultaneous detection of amphetamine, methylenedioxyamphetamine (MDA), methylenedioxymetbamphetamine (MDMA) and methylenedioxyethylamphetamine (MDE). THC was found in 104 (12.2%), cocaine in 230 (27%) and 6-MAM in 141 (16.6%) samples. In addition to 6-MAM, morphine was detected in 87 (10.2%) and heroin in 38 samples (4.5%). The concentrations found were in a range 0.009-16.7 ng/mg for THC, 0.037-129.68 ng/mg for cocaine, 0.028-79.82 ng/mg for 6-MAM, 0.045-53.14 ng/mg for heroin and 0.011-7.800 ng/mg for morphine. The statistical distribution of the drug concentrations compared with the self-reported consumption behaviour of the users may possibly lead to a better understanding of the relationship between drug dosage and corresponding concentrations in hair.     

Detection of methadone in human hair by gas chromatography/mass spectrometry

Summary Determination of methadone in human hair by gas chromatography/mass spectrometry was described. Helium as carrier gas, a 30-m bonded phasefused silica DB-1 capillary column and splitless injection at 230°C temperature were used. The concentrations of methadone and its metabolites were measured in addition by radioimmunoassay (RIA). Both methods GC/MS and RIA showed the presence of methadone in human hair.     

Treatments against hair loss may hinder cocaine and metabolites detection

Recently, some of the hair samples that we routinely analyse for drugs of abuse did not produce valid results for cocaine and metabolites. A series of very intense interfering peaks with ion fragments common to cocaine (CO), and benzoylecgonine (BE) were found to cover up the “cocaine” region of the chromatogram. In one of these cases the subject declared he had used a lotion containing Minoxidil in order to prevent hair loss. Starting from this observation we found that the interfering peaks belonged to four different TMS derivatives of Minoxidil. Minoxidil interference was further investigated by applying Tricoxidil^®, a Minoxidil solution, to the hair of CO-free volunteers and to a CO-positive hair strand dipped into Tricoxidil. Hair were analysed before and after treatment. In both cases interfering peaks were absent in the chromatograms of untreated hair and appeared in treated hair. In the CO-positive hair detection of CO, BE and internal standard was completely hindered after treatment with Minoxidil. Attempts to separate interfering peaks from CO and metabolites by modifying the temperature programme failed. None of the hair washing methods tested (methanol; dichloromethane; sodium dodecyl sulphate water solution, 1% w/v followed by methanol; phosphate buffer 0.1 M, pH 6 followed by methanol) succeeded in removing Minoxidil interference. However, a simple solution to partially overcome the problem was to dry up the derivatised extract, reconstitute it in methanol (in order to switch back Minoxidil derivatives to the native molecule), and re-inject it: owing to the higher polarity, underivatised Minoxidil does not interfere any more with the chromatography of CO, at the expense of the disappearance of BE and ecgonine methyl ester both producing TMS derivatives. This strategy was applied to four real cases where Minoxidil interference was recognised: in two of these cases CO was detected. The problem of Minoxidil interference on CO detection may be limited to procedures involving trimethylsilylation, which is probably the most commonly adopted derivatisation in laboratories performing hair analysis for drugs of abuse.     

Deposition of cocaine in tissue after lethal and repeated sublethal administration to sheep

Summary The deposition of cocaine in tissue after lethal administration to sheep was investigated. In addition, the presence of cocaine in tissue obtained from sheep treated for 30 days with a sublethal dose (2.4 mg/kg b.wt.) after 1-day, 1-week, or 1-month withdrawal was studied. The determination of cocaine was performed by radioimmunoassay. The concentrations measured represent the sum of cocaine and its metabolites. The presence of cocaine was also qualitatively proven by gas chromatography/mass spectrometry. After a lethal dose administration cocaine was found in all investigated organs. The highest concentrations were present in liver, bile, and kidney. In tissue obtained from the sheep treated daily with a sublethal dose and killed after 1-week withdrawal, the concentrations found were significantly lower. After 1-month withdrawal, cocaine was not to be discovered in tissue.Diese Arbeit ist Herrn Prof. Dr. G. Dotzauer zu seinem 75. Geburtstag gewidmet     

毛发移植进展

目的:综述毛发移植的研究进展。方法:采用文献回顾和综合分析方法。结果:毛发移植有许多新的术式、新设备出现,毛发移植的基础研究已达到分子和基因水平,然而毛囊干细胞的研究和毛囊体外培养的临床应用还很不成熟。结论:毛发组织工程学的重大突破将提供更好的毛发移植技术,对整形科技极具现实意义。     

Hair testing for cocaine in living men: an epidemiological study on hair samples collected for various medico-legal purposes

The collection and analysis of hair samples offers higher chances of identifying people addicted to drugs than urine samples. In our study we propose hair analysis as a useful epidemiological indicator to identify the extent of cocaine use in different medico-legal situations: currently the epidemiological trends of substance abuse are estimated indirectly, mainly through large-scale surveys or through medical and crime statistics that may be too generic. We conducted a cross-sectional study, analysing hair samples, collected for various medico-legal purposes, from living men belonging to different categories, in order to complete a previous study conducted in 2016. Of 104 samples collected for different purposes, 55 were positive for drugs (52.88% of total) and, in this group, positivity for cocaine was found in 28 cases (26.92% of total), with a mean concentration of 4.58 ng/mg of hair, and a mean concentration of 0.75 ng/mg of hair of its major metabolite (benzoylecgonine). This datum is in line with the Italian Superior Health Institute reports, which declared a reduction in the use of cannabis and benzodiazepines, but a notable increase in cocaine use. Despite its limitations, this study adds further information to the previous one conducted in the same context, and furnishes an interesting picture of the phenomenon of cocaine addiction in Italy.     

A comparison of 3H-cocaine binding on melanin granules and human hair in vitro

The in vitro experiments on the interaction of ³H-cocaine and melanin from Sepia officinalis confirmed the existence of drug binding sites on melanin granules. The results suggested that the binding of ³H-cocaine to melanin could be analyzed by assuming that the binding to the surface of pigment granules is analogous to the adsorption of a drug on a solid and follows Langmuir adsorption isotherm type I. Scatchard analysis indicated heterogeneity of binding sites. Structural and chemical alterations caused by isolation of the melanoproteins, which are heterogenous in nature and show different physicochemical properties, are considered to be most crucial. The studies on hair samples confirmed that melanin-drug interactions occur on the surface of melanin granules. These seem to be of minor importance compared to the drug-melanoprotein loading during melanogenesis for the observed influence of pigmentation on the drug content of hair fibers. From the results it was concluded that in vitro studies on melanin provide limited information and even drug-soaked hair must be regarded as inappropriate for the study of melanin-drug-binding in hair. Received: 21 August 1996 / Received in revised form: 4 December 1996     

DIA3 phenotyping in human tissues,dental pulps,and hair roots by isoelectric focusing

Summary The polymorphism of DIA3 was investigated in tissues of various human organs, dental pulps, and hair roots by isoelectric focusing. DIA3 types were demonstrated from tissues of brain, prostate, testis, ovary, and uterus, but not from tissues of spleen, pancreas, heart, liver, muscle, lung, skin, and kidney. Determination was possible from dental pulps stored at room temperature for up to 2 weeks and from fresh hair roots. The results show that the DIA3 typing by isoelectric focusing is useful for medicolegal individualization of brain, reproductive organs, teeth, and hairs.     

Myocardial necrosis and cocaine

A quantification of different forms of acute myocardial necrosis, myocardial leukocytic infiltrates and myocardial fibrosis was accomplished in 26 chronic cocaine abusers who died of cocaine intoxication and compared to 45 normal subjects who died from head trauma and 38 who died of acquired immunodeficiency syndrome. The findings were: absence of infarct necrosis, a similar frequency and extent of coagulative myocytolysis (contraction band necrosis) and leukocytic infiltrates in cocaine abusers and normal controls, and an absence of myocardial fibrosis in cocaine abusers. These findings question both the acute and chronic cardiotoxicity of cocaine. The infarct-like pattern in some predisposed subjects may be due to an excess of catecholamine release induced by the drug resulting in coagulative myocytolysis and platelet thrombi. Received: 5 August 1996 / Received in revised form: 3 February 1997     

Evaluation of isotopically labeled internal standards and methods of derivatization for quantitative determination of cocaine and related compounds

Gas chromatography-mass spectrometry (GCMS) is the preferred method for the analysis of drugs/metabolites in biological specimens with use of isotopically labeled analogs of the analytes as internal standards (ISs). An important aspect of the chemical derivatization (CD) for GC-MS analysis is that the CD products derived from the analyte and the selected IS must generate ions suitable for designating the analyte and the IS. These ions should not have significant cross contribution (CC), i.e., IS contribution to the intensities of the ions designated for the analyte, and vice versa. With this in mind, the authors have conducted a search of isotopically labeled analogs of commonly abused cocaine and related compounds (cocaine, norcocaine, benzoylecgonine, cocaethylene, ecgonine, ecgonine methyl ester, anhydroecgonine methyl ester) that are commercially available. These ISs and analytes were derivatized with various groups of reagents, and the CD products were analyzed by GC-MS. MS data are presented in two forms: (1) systematic presentation of fullscan spectra; and (2) tabulation of CC data for ions with potential for designating the ISs and analytes. Many (if not most) of these full-scan spectra are not yet available in the open literature and should be of routine reference value to forensic and clinical laboratories that are engaged in the analysis of these drugs/metabolites. Fullscan MS data were further used to select ion pairs with potential for designating the analytes and ISs in quantitative analysis protocols. The CC data of these ion pairs were evaluated using data collected under the selected ion-monitoring mode and summarized in table format. The data exhibited similar CC characteristics in each alkyl, acetyl, or TMS series. Among the potentially usable ion pairs derived from a specific CD group, there was a trend that the ion pairs with higher mass showed better CC data. The CC data derived from the use of ISs labeled with more deuterium atoms were generally more favorable. These data should save enormous amounts of time and effort for practicing laboratories in their search for optimal analytical parameters.     

Recurrent encephalic hemorrhage associated with cocaine abuse

We report a case of recurrent intracerebral hemorrhage secondary to cocaine abuse in a patient with no other predisposing factors. The hemorrhages were located both supra- and infratentorially. Correspondence to: J. M. Pumar Cebreiro     

Polymorphism of EsD by isoelectric focusing: Phenotyping in human tissues,dental pulps,hair roots,and semen

Summary The polymorphism of EsD was investigated in tissues of various human organs, dental pulps, hair roots, and seminal stains by isoelectric focusing. The method yielded an excellent resolution of the isoenzyme components. The time limits of determination were: in organ tissues 3 weeks, in dental pulps 1 week, and in hair roots several days. The 7-1 type was less stable than the common types. Phenotyping was possible from fresh semen samples, but was unsuccessful from dried seminal stains after storage. The results show that the EsD typing by isoelectric focusing is of practical use for medicolegal individualization of organs, teeth, and hairs.     

Immunochemical studies on blood group A substance from human hair

Summary Blood group A-active substance was extracted from urea-treated human hair with methanol-ethyl ether 1:1, v/v) or chloroform-methanol (1:1, v/v). The serological activity of blood group A substance in the hair was destroyed by A-decomposing enzyme fromClostridium tertium with concomitant development of blood group H activity. It is concluded therefore that the extract from the hair of group A contained blood group A-active glycolipid withN-acetylgalactosamine as the non-reducing sugar.

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Zusammenfassung Das A-blutgruppenaktive Glycolipoid wurde mit Methanol-Äther (1:1, v/v) oder Chloroform-Methanol (1:1, v/v) aus Harnstoff-behandelten Haaren extrahiert. Durch Einwirkung von A-zerstörendem Enzym ausClostridium tertium auf das A-Glycolipoid aus Haaren, kommt nunmehr die H-Eigenschaft nach Inaktivierung von A zum Vorschein. Darum kann essein, daßN-Acetyl-Galactosamin für die A-Aktivität vom Haarglycolipoid benötigt wird.

     

Aortic dissection and cocaine use

Most of the cocaine - deaths are said to be related to cardiovascular complications. This paper addresses a rather infrequent complication of chronic cocaine use, represented by the aortic dissection. The case in point pertains to a 45-year-old, caucasian male, substance abuser who suffered an aortic dissection following the use of cocaine. Blood concentrations of cocaine and benzoylecgonine were considered not to be within a potentially toxic range.     

A body-packer with a cocaine bag stuck in the stomach

Management of patients carrying packets of drugs in the digestive tract is a frequent medical problem.Wereport on a patient who was referred by the police after ingestion of packets of cocaine.After spontaneous elimination of 81 drug packets,the patient had three unremarkable stools.A plain abdominal X-ray disclosed no residual packet but computed tomography(CT) scan showed one in the stomach.As this was not eliminated during the 10 d following ingestion,it was removed through gastrotomy.This case stresses the usefulness of the CT scan to ensure that no residual packet is present before hospital discharge.     

Oesophageal and gastric obstruction in a cocaine body packer

While the management of asymptomatic body packers is mainly conservative, few individuals will require surgery for acute toxicity related to packets rupture, intestinal obstruction or very slow progression of the packages. Obstruction of the lower oesophagus or stomach is not frequently reported. We report the case of a 49-year-old woman who had ingested 92 cocaine-containing packages. She was admitted to the hospital after opioid syndrome related to the intake of morphine and codeine to decrease intestinal transit. The presence of more than 80 packages was suspected in the stomach on the initial abdomen computed tomography. Due to the absence of progression of the packages after four days of well-conducted laxative therapy and to major gastric distension at abdomen tomography, surgery was decided and gastrotomy allowed the evacuation of 80 packages that were still present in the stomach or in the lower oesophagus. In addition, 12 other packages had been retrieved either after laxative therapy (9) or by evacuation via the anal canal (3) after palpation of the intestine during the surgical procedure. No complication was observed.