Y-chromosome haplotypes in Italy: the GEFI collaborative database.

A sample of 1176 males from 10 Italian regions have been typed for DYS19, DYS389-I, DYS389-II, DYS390, DYS391, DYS392, DYS393, and DYS385. Individual haplotype data are available on line. A low degree of variation is present among regions. Use of this database is specifically recommended for forensic applications in Italy.

High sensitivity simultaneous determination in hair of the major constituents of ecstasy (3,4-methylenedioxymethamphetamine, 3,4-methylenedioxyamphetamine and 3,4-methylene-dioxyethylamphetamine) by high-performance liquid chromatography with direct fluorescence detection.

A simple, but sensitive and specific high-performance liquid chromatographic assay for the simultaneous determination of the major constituents of "ecstasy" [i.e. 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxyethylamphetamine (MDE)] with direct fluorimetric detection, particularly intended for the routine analysis of hair, is described. Hair samples (100 mg) were overnight incubated in 1 ml of 0.25 M HCl at 45 degrees C and extracted with a commercial liquid-liquid method. The dried residue reconstituted with 500 microl of 0.05 M NaH2PO4 pH 5.2 was injected. Isocratic reversed-phase liquid chromatography was carried out on a column (250x4.6 mm I.D.) packed with spherical 5-microm poly(styrene-divinylbenzene) particles; the mobile phase was composed of 0.1 M potassium phosphate (pH 3)-acetonitrile (82:18). The excitation and the emission wavelengths were set to 285 and 320 nm, respectively. Under the described conditions, MDA, MDMA and MDE eluted in symmetric peaks with an analysis time of 30 min. The limit of detection was lower than 1 ng/ml, with a signal-to-noise ratio of 5, for each compound in solution, allowing a cut-off of 0.1 ng/mg in the hair matrix to be established. The intra-day precision (n = 6) of the assay was characterised by RSDs between 1.0 and 3.0% and between 0.52 and 0.88% for concentrations of 10 and 100 ng/ml, respectively; in day-to-day precision tests (n = 6), RSDs ranged between 5.12 and 11.12%, respectively, for the same concentrations. Interferences from as many as 92 therapeutic and/or abused drugs currently in use in the population were excluded, including N-methyl-1-(3,4-methylenedioxyphenyl)-2 butanamine (MBDB).

Death from heroin overdose: findings from hair analysis.

BACKGROUND: Morphine analysis of hair is used in forensic toxicology to study the addiction history of heroin addicts. To clarify the features underlying fatal heroin intake, we measured hair morphine content in a group of deceased heroin addicts, to verify a possible correlation between fatal heroin overdoses and the addiction behaviour of these individuals before death. METHODS: 91 deaths were attributed to heroin overdose in Verona, Italy, in 1993-96. We analysed the hair of 37 of these individuals, and of 37 active heroin addicts, 37 former heroin users abstinent from the drug for several months, and 20 individuals with no evidence of exposure to opioids. From each individual, a hair sample of about 150 mg was analysed by RIA and high-performance liquid chromatography, to measure the morphine content. FINDINGS: The mean morphine content in the hair of the addicts who had died was 1.15 ng/mg (SD 2.35 ng/mg; range 0-12.25 ng/mg) compared with 6.07 ng/mg (4.29; 1.15-17.0) in the active heroin addicts, 0.74 ng/mg (0.93; 0.10-3.32) in the abstinent former addicts, and values below the detection limit in the non-exposed group. Hair morphine content among those who had died was significantly lower than that in active heroin consumers (p<.00001), but not significantly different from that in the former addicts (p=0.978). INTERPRETATION: Although our findings may be subject to selection bias, since suitable hair samples were available for only 37 of the 91 addicts who had died, these findings support the theory of high susceptibility to opioid overdose after periods of intentional or unintentional abstinence, due to loss of tolerance. Medical staff running detoxification programmes should be aware of the risk inherent in relapse to heroin after a period of abstinence. Moreover, occasional heroin use without a build-up of tolerance could also give a high risk of overdose.

Hair analysis, a novel tool in forensic and biomedical sciences: new chromatographic and electrophoretic/electrokinetic analytical strategies.

Hair analysis for abused drugs is recognized as a powerful tool to investigate exposure of subjects to these substances. In fact, drugs permeate the hair matrix at the root level and above. Evidence of their presence remains incorporated into the hair stalk for the entire life of this structure. Most abusive drugs (e.g. opiates, cocaine, amphetamines, cannabinoids etc.) and several therapeutic drugs (e.g. antibiotics, theophylline, beta 2-agonists, etc.) have been demonstrated to be detectable in the hair of chronic users. Hence, hair analysis has been proposed to investigate drug abuses for epidemiological, clinical, administrative and forensic purposes, such as in questions of drug-related fatalities and revocation of driving licences, alleged drug addiction or drug abstinence in criminal or civil cases and for the follow-up of detoxication treatments. However, analytical and interpretative problems still remain and these limit the acceptance of this methodology, especially when the results from hair analysis represent a single piece of evidence and can not be supported by concurrent data. The present paper presents an updated review (with 102 references) of the modern techniques for hair analysis, including screening methods (e.g. immunoassays) and more sophisticated methodologies adopted for results confirmation and/or for research purposes, with special emphasis on gas chromatography-mass spectrometry, liquid chromatography and capillary electrophoresis.

Integrated use of hair analysis to investigate the physical fitness to obtain the driving licence: a casework study.

According to the laws presently in force in Italy and the guidelines of the Driving Licence Enforcement Commission of Verona, applicants for the driving licence with a history of drug abuse undergo a medical examination, during which complete anamnestic and clinical data are recorded. On this occasion, a hair sample (50-200 mg) is collected and a urinalysis program is started consisting of EMIT controls for opiates, methadone, cocaine, barbiturates, amphetamines, cannabinoids, benzodiazepines and alcohol carried out on eight seriate samples, collected at random over about 40 days under direct supervision. The positive results from urine immunoassays are confirmed by standardized GC/MS methods. The hair samples are screened for morphine and cocaine, the most abused illicit substances in our region, using commercial RIAs adopting cut-off levels of 0.1 ng/mg. All positive samples and about 10% of negative are confirmed by HPLC. In case of confirmed positive results, the applicant is informed: if the subject denies use of opiates or cocaine in the recent months, he or she has the chance of submitting for analysis a new hair sample, which is analyzed in parallel with the hair remaining from the previous assay. In case of persisting denial, claiming analytical interferences by other drugs or endogenous substances, further confirmation of results can be carried out by CE and/or by qualitative MS/MS. In addition, hair sampling from multiple sites (scalp, axillary, pubic hair) with different susceptibility to contamination from the external sources can be carried out to rule out the possibility of passive contamination. At present, we investigate more than 700 subjects per year. The results of this integrated diagnostic strategy are presented and discussed.

[Sudden infant death syndrome (SIDS) update: etiopathogenesis, epidemiology, legal medicine aspects, cases]. / Morte improvvisa del lattante (SIDS) update: eziopatogenesi, epidemiologia, aspetti medico legali, casistica.

Sudden infant death syndrome (SIDS) is the most common cause of postneonatal infant death in developed countries. The causes of SIDS remain unknown. The principal hypothesis appears abnormality of cardiorespiratory control, sleep-wake regulation. Also in our personal date the autopsy have not been of sufficient specificity and sensitivity to explain the disease.

High-performance liquid chromatographic determination of levodropropizine in human plasma with fluorometric detection.

The present paper describes a new high-performance liquid chromatographic method with fluorescence detection for the analysis of levodropropizine [S-(-)-3-(4-phenylpiperazin-1-yl)-propane-1,2-diol] (Levotuss), an anti-tussive drug, in human serum and plasma. A reversed-phase separation of levodropropizine was coupled with detection of the native fluorescence of the molecule, using excitation and emission wavelengths of 240 nm and 350 nm respectively. The analytical column was packed with spherical 5 microns poly(styrene-divinylbenzene) particles and the mobile phase was 0.1 M NaH2PO4 pH 3-methanol (70:30, v/v), containing 0.5% (v/v) tetrahydrofuran. For quantitation, p-methoxylevodropropizine was used as the internal standard. Samples of 200 microliters of either serum or plasma were mixed with 200 microliters of 0.1 M Na2HPO4 pH 8.9 and extracted with 5 ml of chloroform-2-propanol (9:1, v/v). The dried residue from the organic extract was redissolved with distilled water and directly injected into the chromatograph. The limit of detection for levodropropizine, in biological matrix, was about 1-2 ng/ml, at a signal-to-noise ratio of 3. The linearity was satisfactory over a range of concentrations from 3 to 1000 ng/ml (r2 = 0.99910); within-day precision tested in the range 5-100 ng/ml as well as day-to-day reproducibility proved acceptable, with relative standard deviations better than 1% in most cases. Interferences from as many as 91 therapeutic or illicit drugs were excluded.

Reversed-phase high-performance liquid chromatographic determination of cocaine in plasma and human hair with direct fluorimetric detection.

A simple, but sensitive and specific, high-performance liquid chromatographic assay for cocaine with direct fluorimetric detection, particularly intended for the routine analysis of hair and blood samples, is described. Benzoylecgonine, eluting before cocaine in a completely resolved peak, is also detectable. Detection is based on the weak native fluorescence of cocaine and benzoylecgonine, depending on the benzene ring present in both molecules. Hair samples (20-200 mg) were incubated overnight in 2 ml of 0.25 M HCl at 45 degrees C and extracted with a commercial liquid-liquid method; the dried residue reconstituted with 500 microliters of 0.05 M NaH2 PO4 (PH 5.2) was injected. Blood plasma samples (200 microliters) were mixed with 150 microliters of 0.1 M Na2 HPO4 (pH 8.9) and extracted with 5 ml of chloroform-2-propanol (9:1); the organic phase was evaporated and the residue dissolved and injected as above. Isocratic reversed-phase liquid chromatography was carried out on a column (150 x 4.6 mm I.D.) packed with spherical 5-microns poly(styrene-divinylbenzene) particles; the mobile phase was 0.1 M potassium phosphate (pH 3)-methanol-tetrahydrofuran (70:25:5). The excitation and emission wavelengths were set at 230 and 315 nm, respectively. Under the described conditions, cocaine eluted in a symmetrical peak with a capacity factor of about 5. The limit of detection was about 1 ng/ml (0.2 ng injected), with a signal-to-noise ratio of 3.(ABSTRACT TRUNCATED AT 250 WORDS)