State of the art in hair analysis for detection of drug and alcohol abuse.

http://www.x-pertise.com/x-pertise.com/News_files/SoHT%20Guidelines%20FSI%20online%2016NOV11.pdf

Society of Hair Testing guidelines for drug testing in hair

EASL Clinical Practice Guidelines: Management of alcohol-related liver disease

The vast, diverse universe of organic pollutants is a formidable challenge for environmental sciences, engineering, and regulation. Nontarget screening (NTS) based on high resolution mass spectrometry (HRMS) has enormous potential to help characterize this universe, but is it ready to go for real world applications? In this Feature article we argue that development of mass spectrometers with increasingly high resolution and novel couplings to both liquid and gas chromatography, combined with the integration of high performance computing, have significantly widened our analytical window and have enabled increasingly sophisticated data processing strategies, indicating a bright future for NTS. NTS has great potential for treatment assessment and pollutant prioritization within regulatory applications, as highlighted here by the case of real-time pollutant monitoring on the River Rhine. We discuss challenges for the future, including the transition from research toward solution-centered and robust, harmonize...

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Nontarget Screening with High Resolution Mass Spectrometry in the Environment: Ready to Go?

Solid-phase microextraction in biomedical analysis.

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 40–4347 pg/mg (average: 628, median 374 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

Ethyl glucuronide in hair: Is it a reliable marker of chronic high levels of alcohol consumption?

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.

Determination of ethyl glucuronide in hair samples by liquid chromatography/electrospray tandem mass spectrometry.

Aims This study aims to define a cut-off concentration for ethyl glucuronide in hair to determine if there was a history of heavy drinking.



Settings  Pavia, Italy.



Participants  We analysed hair samples from 98 volunteers among teetotallers, social drinkers and heavy drinkers, whose ethanol daily intake (EDI) was estimated by means of a written questionnaire.



Measurements  Ethyl glucuronide hair concentration (HEtG) was measured by liquid chromatography-tandem mass spectrometry (lower limit of quantification: 3 pg/mg) using a fully validated method.



Findings  The HEtG level providing the best compromise between sensitivity (0.92) and specificity (0.96) at detecting an EDI of 60 g or higher during the last 3 months was 27 pg/mg. None of the factors examined among those known to affect ethanol metabolism and/or the diagnostic power of other markers of ethanol use or hair analyses, including age, gender, body mass index, tobacco smoke, prevalent beverage, hair colour, cosmetic treatments and hygienic habits was found to influence marker performance significantly. However, the slight differences in HEtG performance observed for some factors (e.g. body mass index, smoke and hair treatments) require further studies on larger groups of individuals in order to assess their influence more precisely.



Conclusions  Our results confirm further that HEtG is a sensitive and specific marker of chronic heavy drinking.

Ethyl glucuronide in hair. A sensitive and specific marker of chronic heavy drinking.

Effect of bleaching on ethyl glucuronide in hair: An in vitro experiment

Electrospray ionization (ESI)-time-of-flight (TOF) MS enables searching a wide number of pharmaco/toxicologically relevant compounds (PTRC) in biosamples. However, the number of identifiable PTRC depends on extension of reference database of chemical formulas/compound names. Previous approaches proposed in-house or commercial databases with limitations either in PTRC number or content (e.g., few metabolites, presence of non-PTRC). In the frame of development of a ESI-TOF PTRC screening procedure, a subset of PubChem Compound as reference database is proposed. Features of this database (approximately 50,500 compounds) are illustrated, and its performance evaluated through analysis by capillary electrophoresis (CE)-ESI-TOF of hair/blood/urine collected from subjects under treatment with known drugs or by comparison with reference standards. The database is rich in parent compounds of pharmaceutical and illicit drugs, pesticides, and poisons and contains many metabolites (including about 6000 phase I metabolites and 180 glucuronides) and related substances (e.g., impurities, esters). The average number of hits with identical chemical formula is 1.82 +/- 2.27 (median = 1, range 1-39). Minor deficiencies, redundancies, and errors have been detected that do not limit the potential of the database in identifying unknown PTRC. The database allows a much broader search for PTRC than other commercial/in-house databases of chemical formulas/compound names previously proposed. However, the probability that a search retrieves different PTRC having identical chemical formula is higher than with smaller databases, and additional information (anamnestic/circumstantial data, concomitant presence of parent drug and metabolite, selective sample preparation, liquid chromatographic retention, and CE migration behavior) must be used in order to focus the search more tightly.

Implementation and performance evaluation of a database of chemical formulas for the screening of pharmaco/toxicologically relevant compounds in biological samples using electrospray ionization-time-of-flight mass spectrometry.

Aldo Polettini

Papers

Ethyl glucuronide in hair: Is it a reliable marker of chronic high levels of alcohol consumption?

Determination of ethyl glucuronide in hair samples by liquid chromatography/electrospray tandem mass spectrometry.

Ethyl glucuronide in hair. A sensitive and specific marker of chronic heavy drinking.

Effect of bleaching on ethyl glucuronide in hair: An in vitro experiment

Implementation and performance evaluation of a database of chemical formulas for the screening of pharmaco/toxicologically relevant compounds in biological samples using electrospray ionization-time-of-flight mass spectrometry.