Centro Regionale Antidoping A Bertinaria

Orbassano, Italy

Centro Regionale Antidoping A Bertinaria

Orbassano, Italy
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Salomone A.,Centro Regionale Antidoping A Bertinaria | Gerace E.,Centro Regionale Antidoping A Bertinaria | Gerace E.,University of Turin | Brizio P.,Centro Regionale Antidoping A Bertinaria | And 4 more authors.
Journal of Pharmaceutical and Biomedical Analysis | Year: 2011

A fast liquid chromatographic/tandem mass spectrometric method was developed for the simultaneous determination in human urine of seventeen benzodiazepines, four relevant metabolites together plus zolpidem and zopiclone. The sample preparation, optimized to take into account the matrix effect, was based on enzymatic hydrolysis and liquid-liquid extraction. The separation of the twenty-three analytes was achieved in less than eight minutes.The whole methodology was fully validated according to UNI EN ISO/IEC 17025:2005 rules and 2006 SOFT/AAFS guidelines. Selectivity, linearity range, identification (LOD) and quantitation (LOQ) limits, precision, accuracy and recovery were evaluated. For all the species the signal/concentration linearity was satisfactory in the 50-1000 ng/mL concentration range. The limits of detection ranged from 0.5 to 30 ng/mL and LOQs from 1.7 to 100.0. ng/mL. Precisions were in the ranges 5.0-11.8%, 1.5-11.0% and 1.1-4.4% for low (100 ng/mL), medium (300 ng/mL) and high (1000 ng/mL) concentration, respectively. The accuracy, expressed as bias% was within ±25% for all the analytes. The recovery values, evaluated at 300 ng/mL concentration, ranged from 56.2% to 98.8%. The present method for the determination of several benzodiazepines, zolpidem and zopiclone in human urine proved to be simple, fast, specific and sensitive. The quantification by LC-MS/MS was successfully applied to 329 forensic cases among driving re-licensing, car accidents and alleged sexual violence cases. © 2011 Elsevier B.V.

Gerace E.,University of Turin | Gerace E.,Centro Regionale Antidoping A Bertinaria | Salomone A.,Centro Regionale Antidoping A Bertinaria | Fasano F.,University of Turin | And 5 more authors.
Analytical and Bioanalytical Chemistry | Year: 2011

Selective androgen receptor modulators (SARMs) represent an emerging class of drugs likely to be abused in sport. For clinical applications, these substances provide a promising alternative to testosterone-replacement therapies and their advantages include oral bioavailability, androgen receptor specificity, tissue selectivity, and the absence of steroid-related side effects. Although not yet commercially available, since January 2008 SARMs have been included on the prohibited list issued yearly by the World Anti-Doping Agency (WADA), so control laboratories need to update their procedures to detect either the parent drugs or their metabolites. Within this context, two quinolinone SARM models were synthesized and automatically characterized to update the existing routine screening procedures. The conditions for the new target analytes are compatible with the existing laboratory protocols used for both in-competition and out-of-competition controls and can be included in them. Validation parameters according to ISO 17025 and WADA guidelines were successfully determined. For analytical determinations, spiked urine samples were hydrolyzed and extracted at pH 9.6 with 10 mL of tert-butyl methyl ether. Then, the analytes were subsequently converted into trimethylsilyl derivatives and detected by gas chromatography-mass spectrometry. The absence of interferents, together with excellent repeatability of both retention times and the relative abundances of diagnostic ions, allowed proper identification of all SARM analytes. The analytes' quantification was linear up to 500 ng/mL and precision criteria were satisfied (coefficient of variation less than 25% at 10 ng/mL). The limits of detection were 1 ng/mL for both SARMs, whereas recovery values were between 95.5 and 99.3%. The validated method can be efficiently used for urine screening of the 2-quinolinone-derived SARMs tested. [Figure not available: see fulltext.] © 2010 Springer-Verlag.

Salomone A.,Centro Regionale Antidoping A Bertinaria | Di Corcia D.,Centro Regionale Antidoping A Bertinaria | Gerace E.,Centro Regionale Antidoping A Bertinaria | Vincenti M.,Centro Regionale Antidoping A Bertinaria | Vincenti M.,University of Turin
Journal of Analytical Toxicology | Year: 2011

Mirtazapine, escitalopram, and valproic acid are newer antidepressant drugs than traditional tricyclic antidepressants and are supposed to be less toxic. Nevertheless, intoxication cases due to their overdosage have been repeatedly reported. In the case presently reported, a 64-year-old woman with a previous history of chronic depression was found dead in her apartment. Several packages of pharmaceutical drugs were found, including mirtazapine, escitalopram, and valproic acid. During the autopsy, no evidence of natural disease or trauma was found to account for this death. In order to determine whether massive drug assumption might have determined a lethal intoxication, heart blood, urine, and gastric content were collected and submitted to toxicological analysis. Specific liquid chromatography-tandem mass spectrometry protocols were purposely developed and validated. Blood concentrations of mirtazapine, escitalopram, and valproic acid were 20.3, 65.5, and 417 mg/L, respectively, whereas urine concentrations were 17.0, 94.5, and 423 mg/L, respectively. High concentrations of these drugs were also detected in the gastric content, confirming their ingestion shortly before death. The agreement between authoptic examination by forensic pathologists and toxicological findings are consistent with the suicidal hypothesis, where the death arose by drug intoxication due to simultaneous high-dosage ingestion of mirtazapine, escitalopram, and valproic acid.

Vincenti M.,Centro Regionale Antidoping A Bertinaria | Vincenti M.,University of Turin | Leporati M.,University of Turin | Capra P.,Instituto Zooprofilattico Sperimentale del Piemonte | And 4 more authors.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2012

Prednisolone is a synthetic glucocorticoid widely employed in bovine clinical practice that may also be used illegally as a growth promoter. Recent in vitro and in vivo studies lend support to the hypothesis that prednisolone could be synthesised from cortisol in untreated cattle subjected to stressful events. To verify such a hypothesis, a field survey was conducted on urine samples collected from 131 guaranteed untreated cows and analysed for the presence of prednisolone and prednisone - in some instances also for cortisol and cortisone - with a validated LC/MS-MS method. None of the examined samples exhibited either prednisolone levels higher than the CCα limit (around 0.70 μg l-1) or prednisone, being therefore officially compliant for both analytes. Trace amounts of prednisolone, approximately estimated in the range 0.1-0.3 μg l-1 were found in only seven samples from cows also showing urinary cortisol and cortisone levels higher than those detected in negative specimens, as the result of a probable stress condition. © 2012 Copyright Taylor and Francis Group, LLC.

Gerace E.,Centro Regionale Antidoping A Bertinaria | Salomone A.,Centro Regionale Antidoping A Bertinaria | Abbadessa G.,University of Turin | Racca S.,University of Turin | And 2 more authors.
Journal of Pharmaceutical Analysis | Year: 2012

A fast screening protocol was developed for the simultaneous determination of nine anti-estrogenic agents (aminoglutethimide, anastrozole, clomiphene, drostanolone, formestane, letrozole, mesterolone, tamoxifen, testolactone) plus five of their metabolites in human urine. After an enzymatic hydrolysis, these compounds can be extracted simultaneously from urine with a simple liquidliquid extraction at alkaline conditions. The analytes were subsequently analyzed by fast-gas chromatography/mass spectrometry (fast-GC/MS) after derivatization. The use of a short column, high-flow carrier gas velocity and fast temperature ramping produced an efficient separation of all analytes in about 4 min, allowing a processing rate of 10 samples/h. The present analytical method was validated according to UNI EN ISO/IEC 17025 guidelines for qualitative methods. The range of investigated parameters included the limit of detection, selectivity, linearity, repeatability, robustness and extraction efficiency. High MS-sampling rate, using a benchtop quadrupole mass analyzer, resulted in accurate peak shape definition under both scan and selected ion monitoring modes, and high sensitivity in the latter mode. Therefore, the performances of the method are comparable to the ones obtainable from traditional GC/MS analysis. The method was successfully tested on real samples arising from clinical treatments of hospitalized patients and could profitably be used for clinical studies on anti-estrogenic drug administration. © 2012 Xian Jiaotong University. Productionand hosting by Elsevier B.V. All rights reserved.

Gerace E.,Centro Regionale Antidoping A Bertinaria | Gerace E.,University of Turin | Salomone A.,University of Turin | Pellegrino S.,Centro Regionale Antidoping A Bertinaria | And 2 more authors.
Forensic Science International | Year: 2012

A case of intoxication by haloperidol is reported. Haloperidol is a butyrophenone derivative commonly used in many hospital units as an antipsychotic agent. Adverse reactions due to haloperidol intoxication include drowsiness, blurred vision, extrapyramidal effects, tardive dyskinesia, tachycardia, hypotension and muscular rigidity. In August 2008, a 49 year-old female nurse started feeling various symptoms such as muscular rigidity, drowsiness and buccal dyskinesia. After 3 months, she was hospitalized for the worsening of these symptoms. Four months later, she showed once more the same symptoms. Two open water bottles from which the nurse used to drink in the hospital were confiscated and analyzed. Moreover, the nurse was asked to give a sample of her hair for executing the inherent toxicological analyses. Haloperidol was found in both bottles 1 and 2 at a concentration of 31.5 μg/mL and 43.6 μg/mL, respectively. Based on segmental hair analysis, it was deduced that the nurse consumed haloperidol in the approximate period from August 2008 to March 2009. The higher levels of haloperidol in hair were found in accordance with the periods of most severe appearance of symptoms, requiring the hospitalization of the nurse. The analysis of preservatives and excipients led us to conclude that the pharmaceutical drug was probably added to the water bottles as Haldol 2. mg/mL oral solution. © 2011 Elsevier Ireland Ltd.

Salomone A.,Centro Regionale Antidoping A Bertinaria | Gerace E.,Centro Regionale Antidoping A Bertinaria | Di Corcia D.,Centro Regionale Antidoping A Bertinaria | Martra G.,University of Turin | And 3 more authors.
International Journal of Legal Medicine | Year: 2012

In drug-facilitated crimes, victims are subjected to nonconsensual acts while they are incapacitated by the effects of a drug. A specific LC-MS/MS protocol for determining benzodiazepines and hypnotics at low concentration in hair specimens was developed and validated in order to target the allegedly administered drugs on a chronological basis. In the case hereby reported, a 26- year-old woman claimed to have been sexually assaulted after being administered an allegedly drugged coffee, but toxicological analysis of urine and blood provided no evidence of any drug intake. Subsequently, a second woman accused the same man of sexual abuse. Hence, the suspect was prosecuted. Specimens were collected from four subjects (two alleged victims, the suspect and his wife) and segmental hair analysis was performed. The results revealed that zolpidem was present at low picogram per milligram concentration in three out of eleven segments of hair specimen obtained from the first of the alleged victims, offering plain evidence of single or sporadic exposure, whereas the agent was detected in the high picogram per milligram range in the hair collected from suspect's wife, coherently with therapeutic administration. The presence of interfering signals typical of the keratin-containing matrix was found and possible hair degradation by cosmetic treatments was investigated by electron microscopy, so as to obtain a judicious interpretation of the analytical findings. © Springer-Verlag 2012.

Salomone A.,Centro Regionale Antidoping A Bertinaria | Luciano C.,Centro Regionale Antidoping A Bertinaria | Di Corcia D.,Centro Regionale Antidoping A Bertinaria | Gerace E.,Centro Regionale Antidoping A Bertinaria | And 2 more authors.
Drug Testing and Analysis | Year: 2014

Among the new psychoactive products, herbal mixtures containing synthetic cannabimimetics are likely the most abused worldwide. In this study, a specific ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the detection of 23 synthetic cannabinoids in hair samples was developed in order to (1) expand the number of screened compounds, coherent with new substances emerging in the European territory, (2) evaluate their consumption on a large period of examination, and (3) evaluate the diffusion of cannabimimetics among different populations of drug consumers. The method employs digestion of hair sample with NaOH followed by extraction with n-hexane/ethylacetate, and injection into the UHPLC-MS/MS system. After validation, the method was applied to the analysis of 344 hair samples previously tested in our laboratory for the most common drugs. Overall, 15 samples were found positive for at least one synthetic cannabinoid. Coherent with previously published results, the present data show that young males, former or still active Cannabis consumers, represent the population most often involved in synthetic cannabimimetics consumption. Several cases of poly-abuse were also determined. The drug most frequently detected was JWH-073 (11 samples) generally at low concentration (mean 7.69 ± 14.4 pg/mg, median 1.9 pg/mg, range 1.6-50.5 pg/mg), followed by JWH-122 (8 samples, mean concentration: 544 ± 968 pg/mg, median 28.4 pg/mg, range 7.4-2800 pg/mg). Other detected drugs included JWH-250, JWH-081, JWH-018, JWH-210, JWH-019, and AM-1220. For several positive samples, the synthetic cannabinoid concentration was lower than 50 pg/mg, underlining the need for established cut-off values for discrimination between chronic consumption and occasional use (or external contamination). © 2013 John Wiley & Sons, Ltd.

PubMed | Labor Krone, X Pertise Consulting, Centro Regionale Antidoping A Bertinaria, a Pentwyn Business Center and University of Zürich
Type: Journal Article | Journal: Drug testing and analysis | Year: 2016

Guidelines for Legally Defensible Workplace Drug Testing have been prepared and updated by the European Workplace Drug Testing Society (EWDTS). They are based on the 2010 version published by Pascal Kintz and Ronald Agius (Guidelines for European workplace drug and alcohol testing in hair. Drug Test. Anal. 2010, 2, 367) and in concordance with the Society of Hair Testing guidelines (Society of Hair Testing guidelines for drug testing in hair. Forensic Sci. Int. 2012, 218, 20-24). The European Guidelines are designed to establish best practice procedures whilst allowing individual countries to operate within the requirements of national customs and legislation. The EWDTS recommends that all European laboratories that undertake legally defensible workplace drug testing use these guidelines as a template for accreditation. Copyright 2016 John Wiley & Sons, Ltd.

A simple procedure for the quantitative detection of JWH-018, JWH-073, JWH 200, JWH-250, HU-210, Δ(9)-tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN) in hair has been developed and fully validated. After digestion with NaOH and liquid-liquid extraction, the separation was performed with an ultra-high performance liquid chromatography system coupled to a triple quadrupole mass spectrometer operating in the selected reaction monitoring mode. The absence of matrix interferents, together with excellent repeatability of both retention times and relative abundances of diagnostic transitions, allowed the correct identification of all analytes tested. The method was linear in two different intervals at low and high concentration, with correlation coefficient values between 0.9933 and 0.9991. Quantitation limits ranged from 0.07 pg/mg for JWH-200 up to 18 pg/mg for CBD The present method for the determination of several cannabinoids in hair proved to be simple, fast, specific and sensitive. The method was successfully applied to the analysis of 179 real samples collected from proven consumers of Cannabis, among which 14 were found positive to at least one synthetic cannabinoid. Copyright © 2012 John Wiley & Sons, Ltd.

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