The mysterious death of Francesco I de' Medici and Bianca Cappello: an arsenic murder?
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Arsenic↗
Biomedical subjects
Aldo Polettini
Publications and source records attributed to Aldo Polettini.
Markers of chronic alcohol use in hair: comparison of ethyl glucuronide and cocaethylene in cocaine users.
Two direct ethanol metabolites, namely ethyl glucuronide (EtG) and cocaethylene (CE), in the hair of cocaine (COC) users were compared in this study. Hair samples (n=68) were submitted to the determination of EtG (by liquid chromatography-electrospray-tandem mass spectrometry) and of COC and metabolites, including CE (by gas chromatography-mass spectrometry). Quantitative and qualitative results were compared. No quantitative correlation was found between EtG and CE, as well as between EtG and the cocaethylene concentration divided by the concentration of COC and its metabolites (benzoylecgonine and ecgonine methylester, as COC equivalents). Nevertheless, many factors are supposed to affect the amount of the two substances incorporated in the hair matrix, such as the subject's habits in ethanol and COC use, genetic variability in the metabolism of both substances, and the different chemical and physical properties of EtG and CE. When establishing a cut-off of 4 pg/mg for EtG and of 200 pg/mg for CE, 47 samples tested positive for EtG and 41 samples tested positive for CE; 12 samples out of the 47 EtG-positives tested negative for CE (25%), whereas 6 samples out of the 41 CE-positives tested negative for EtG (15%). According to these data, EtG appears to be a more sensitive and specific marker of non-moderate alcohol users than CE.
Adolescent↗
Ethyl glucuronide and ethyl sulphate determination in serum by liquid chromatography-electrospray tandem mass spectrometry.
BACKGROUND: Ethyl glucuronide (EtG) and ethyl sulphate (EtS) are two ethanol metabolites that can be detected in serum up to 8 h after ethanol elimination. Their presence is therefore indicative of recent ethanol consumption in case of delayed sampling after an event (e.g. car crash). METHODS: A LC-ESI-MS-MS method for the determination of EtG and EtS in serum was developed and validated. Two product ions together with the parent ions were monitored for identification. Pentadeuterated-EtG was used as internal standard. RESULTS: Excellent linearity for EtG (from 0.22 to 45 micromol/l) and EtS (from 0.40 to 80 micromol/l) was observed (r(2)>or=0.9998). LOD and LLOQ were 0.04 and 0.20 micromol/l for EtG and 0.08 and 0.40 micromol/l for EtS, respectively. Accuracy (bias) and precision (relative standard deviation), studied at four different quality control levels, were always better than 7%. Matrix effects were found to be negligible. The method was applied to several samples obtained from known alcoholics and social drinkers. CONCLUSIONS: A sensitive and specific determination of EtG and EtS in serum samples was achieved despite a simple and fast sample preparation. To our knowledge, this is the first fully validated method for the simultaneous determination of the two alcohol metabolites in serum.
Alcohol Drinking↗
Determination of ethyl glucuronide in hair samples by liquid chromatography/electrospray tandem mass spectrometry.
A method for the determination of ethyl glucuronide (EtG) in hair samples, using liquid chromatography/electrospray tandem mass spectrometry (LC/ESI-MS/MS), was developed and validated. The treatment of hair samples was as follows: to 100 mg of washed (dichloromethane followed by methanol, 1 ml each) and cut (1-2 mm) material, 700 microl of water, 20 microl of internal standard solution (pentadeuterated EtG, D(5)-EtG, 500 microg/l) and 20 microl of methanol were added. Samples were incubated at 25 degrees C overnight and then ultrasonicated for 2 h. Finally, 8 microl of the centrifuged solution (13,000 rpm) were analyzed by LC/ESI-MS/MS in negative ion mode. The surviving ions of EtG and D(5)-EtG were monitored together with the following MRM transitions: m/z 221 --> 75, m/z 221 --> 85 (EtG) and m/z 226 --> 75, m/z 226 --> 85 (D(5)-EtG). The method exhibited a mean correlation coefficient better than 0.9998 over the dynamic range (3-2000 pg/mg). The lower limit of quantification (LLOQ) and the limit of detection (LOD) were 3 and 2 pg/mg respectively. The intra- and interday precision and accuracy were studied at four different concentration levels (3, 5, 56 and 160 pg/mg) and were always better than 7% (n = 5). Matrix effects did not exceed 20%. The method was applied to several hair samples taken from autopsies of known alcoholics, from patients in withdrawal treatment, from social drinkers, from adult teetotalers and from children not exposed to ethanol, with EtG concentrations globally ranging from < or =2 to 4180 pg/mg.
Alcoholism↗
Ethyl glucuronide in hair: Is it a reliable marker of chronic high levels of alcohol consumption?
AIMS: This study aims to investigate the relationship between ethanol daily intake (EDI) and the levels of ethyl glucuronide in hair. DESIGN: Ethyl glucuronide concentration was determined in hair samples from different classes of ethanol drinkers and results were compared with the reported information about drinking habits. SETTING: Pavia, Italy. PARTICIPANTS: Twenty-two known alcoholics, 21 volunteers self-reporting an EDI from 2 to 60 g, and seven teetotallers were involved in this study. MEASUREMENTS: Ethyl glucuronide determination in hair samples was performed by liquid chromatography-tandem mass spectrometry (limit of detection: 2 pg/mg, lower limit of quantification: 3 pg/mg). FINDINGS: Current known alcoholics (n = 21) had ethyl glucuronide hair concentration in the range 4.0-434.7 pg/mg (average: 62.8, median 37.4 pg/mg); ethyl glucuronide was not detected in hair samples from teetotallers (n = 7); all volunteers reporting an EDI of at least 30 g ('non-moderate drinkers' according to the US Department of Health and Human Services) tested positive for ethyl glucuronide (cut-off: 4 pg/mg). All volunteers declaring an ethanol daily intake higher than 40 g ('heavy drinkers' according to the World Health Organization, Regional Committee for Europe) tested positive for this compound (cut-off: 5 pg/mg). The application of a cut-off of either 4 pg/mg or 5 pg/mg resulted in one false positive, coming from a volunteer asserting an ethanol daily intake of 30 g. No false negatives were found. CONCLUSIONS: The concentration of ethyl glucuronide in hair appears to correlate with EDI.
Adult↗
The role of cocaine in heroin-related deaths. Hypothesis on the interaction between heroin and cocaine.
In recent years, there has been an increase in cocaine-related deaths at the Department of Legal Medicine and Public Health of Pavia, probably reflecting the rising trend in cocaine use in Western Europe. Deaths from cocaine alone have increased from 6 cases in 1979-1991 (1.5% of drug-of-abuse deaths) to 13 in 1992-2002 (3.2%) and comparing the same periods, heroin-related deaths (HRDs) involving cocaine more than doubled from 8 (1.9%) to 22 (5.4%). In an attempt to investigate the role of cocaine in HRDs, acute narcotic death cases testing positive for cocaine use (blood cocaine or metabolite concentration >0.01 mg/l, COC+) were examined. Only cases from 1997 to 2001 were considered as in this period all data were obtained using the same analytical procedures (free morphine and total morphine by DPC Coat-A-Count radioimmunoassay before and after enzymatic hydrolysis, cocaine and metabolites in blood by SPE, TMS derivatization and gas chromatography-mass spectrometry (GC-MS)). The median, minimum and maximum concentrations of free morphine in blood (FM) and total morphine in blood (TM), urine (UM) and bile (BM) in the COC+ group (n = 9) were compared with those calculated in the group of "pure" HRDs (no other drugs detected in blood, COC-, n = 30). Differences among the medians in the two groups were statistically evaluated using the two-tailed Mann-Whitney U-Test. Statistical analysis was also carried out including in both groups cases with a blood alcohol concentration (BAC) > 20 mg/L (COC+, n = 19; COC-, n = 76). For the COC+ group, median TM was lower (0.32 mg/l versus 0.90 mg/l, P = 0.0214), median FM was lower, but not statistically significant (0.08 mg/l versus 0.28 mg/l, P = 0.1064), FM/TM ratio was similar (0.33 and 0.35), UM was also similar (21.0 mg/l and 18.0 mg/l), and BM was higher (90.0 mg/l versus 49.0 mg/l, P = 0.0268). Similar comparison results were obtained by repeating statistical analyses after including in the two groups cases with positive BAC. The picture observed for HRD cases involving cocaine is very different from what was previously observed for HRD cases involving ethanol [A. Polettini, A. Groppi, M. Montagna, The role of alcohol abuse in the etiology of heroin related deaths: evidence for pharmacokinetic interactions between heroin and alcohol, J. Anal. Toxicol. 23 (1999) 570-576], and updated with more recent data; in the high-ethanol (HE, BAC > 1000 mg/l) group, TM was lower than in the low-ethanol (LE, BAC <or= 1000 mg/l) group (0.59 mg/l versus 0.90 mg/l, P = 0.0180), FM/TM ratio was higher (0.66 versus 0.43, P = 0.0038), FM was equal, UM was lower (0.21 mg/l versus 13.0 mg/l, P = 0.0001), as was BM (10.0 mg/l versus 26.5 mg/l, P = 0.0156). The observed lower TM in the COC+ group supports the hypothesis of an interaction of cocaine with heroin in the cause of death, as previously observed for the heroin-ethanol interaction. However, ethanol results suggested that a pharmacokinetic interaction is prevalent (inhibition of heroin metabolism, as suggested by the increased FM/TM ratio, resulting in reduced urinary and biliary excretion). In the case of cocaine, a pharmacodynamic interaction seems to prevail, as the FM/TM ratio remains unchanged and UM and BM are not lower in the COC+ group. The hypothesis of a pharmacodynamic interaction of cocaine with heroin could not be confirmed owing to the paucity of data and the many uncontrolled variables involved.
Adolescent↗
Direct determination of the ethanol metabolites ethyl glucuronide and ethyl sulfate in urine by liquid chromatography/electrospray tandem mass spectrometry.
A liquid chromatography/electrospray tandem mass spectrometry (LC/ESI-MS/MS) method for the determination in urine samples of two ethanol metabolites, ethyl glucuronide (EtG) and ethyl sulfate (EtS), was developed and validated. Pentadeuterated EtG was used as internal standard for both EtG and EtS. In addition to the surviving ions, two MS/MS reactions were monitored for each analyte, with the deprotonated molecule as precursor ion: m/z 221 --> 75, m/z 221 --> 85 (EtG), and m/z 125 --> 97, m/z 125 --> 80 (EtS). Sample pretreatment, though very simple and rapid (1:50 water dilution and centrifugation of 50 muL of urine), was found to contain the occurrence of matrix effects. The method was accurate and precise over the linear dynamic range (0.05-10 mg/L). The analytes were stable in frozen urine for at least 1 month. The assay was applied to several authentic urine samples from social drinkers and to alcoholic beverages.
Alcoholic Beverages↗
A rapid screening procedure for drugs and poisons in gastric contents by direct injection-HPLC analysis.
A high performance liquid chromatographic method for toxicological drug screening of gastric content has been developed. The samples were diluted (1:3-1:30) in 0.01 N hydrochloric acid and injected into a reverse phase column for separation by gradient elution. Mobile phase consisted of solvent A (acetonitrile/water 90:10, 0.01 M sodium dodecylsulphate, 0.5% v/v glacial acetic acid) and solvent B (water/acetonitrile 90:10, 0.01 M sodium dodecylsulphate, 0.5% v/v glacial acetic acid); the gradient was programmed from 20 to 80% A in 30 min. The flow was kept constant at 1.5 ml/min. Two home-made internal standards, butyrylsalicylic acid and diacetyltubocurarine with retention times of 5.6 and 21.4 min, respectively, were used. Drugs are identified by matching their relative retention times and UV spectra (200-400 nm) with those contained in a home made library of more than 340 reference compounds (9 analgesics, 22 antidepressants, 30 antihistamines, 14 antihypertensives, 21 antirheumatics, 15 beta-blockers, 9 bronchodilators, 10 Ca antagonists, 14 diuretics, 26 neuroleptics, 25 tranquilizers, and other significant xenobiotic compounds). The fluorometric (FLD) emission spectrum (280-700 nm; excitation wavelength, 230 nm) was used as a further identification. At 50mg/l analyte concentrations, the injection of gastric content after dilution (1:3) produced S/N ratios in the range 8-140. The method is simple, rapid, rather inexpensive and proved to be a useful means of investigation if used in combination with GC-MS screening in blood. On the other hand, the system suffers from a relatively limited sensitivity for compounds with a low UV absorption and from interferences due to the presence in the matrix of some highly UV- and FL-responsive compounds (e.g. tryptophan).
Chromatography, High Pressure Liquid↗
Forensic toxicology: web resources.
Forensic toxicology is the study and practice of the application of toxicology to the purposes of the law. The internet provides abundant web-sites and resources for the practicing forensic toxicologist and those interested in the field of forensic toxicology. This review includes a description of web-sites, databases of toxicological and analytical data, and web-based journals, forums and mailing lists.
Databases as Topic↗
Applications of liquid chromatography-mass spectrometry in doping control.
This paper reviews liquid chromatographic-mass spectrometric (LC-MS) procedures for the screening, identification and quantification of doping agents in urine and other biological samples and devoted to drug testing in sports. Reviewed methods published approximately within the last five years and cited in the PubMed database have been divided into groups using the same classification of the 2004 World Anti-Doping Agency (WADA) Prohibited List. Together with procedures specifically developed for anti-doping analysis, LC-MS applications used in other fields (e.g., therapeutic drug monitoring, clinical and forensic toxicology, and detection of drugs illicitly used in livestock production) have been included when considered as potentially extensible to doping control. Information on the reasons for potential abuse by athletes, on the requirements established by WADA for analysis, and on the WADA rules for the interpretation of analytical findings are provided for the different classes of drugs.
Chromatography, Liquid↗