Celbridge, Ireland
Celbridge, Ireland

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Tkachenko A.,U.S. Food and Drug Administration | Clark J.,State Laboratory | Knutson N.,State Laboratory | Wallace B.,New Hampshire State Public Health Laboratory | And 6 more authors.
Food and Chemical Toxicology | Year: 2015

Four LC-MS/MS methods were developed to quantify melamine (MEL) and cyanuric acid (CYA) in various pig tissues at or above the level of concern (2.5 mg/kg). Pigs treated with 200 mg/kg bw/day CYA daily for 7 days did not accumulate significant residue concentrations in muscle, liver or kidney. Pigs treated with 200 mg/kg bw MEL daily for 7 or 28 days had MEL residues in muscles (3-13 ppm), liver (2.8-14.1 ppm) and kidney (9.4-27.2 ppm). Treatment with MEL and CYA at 100 mg/kg bw of each triazine daily for 7 days resulted in MEL (26-59 ppm in muscle, 30-49 ppm in liver and 367-6300 ppm in kidney) and CYA (1.8-5.8 ppm in muscle, 2.6-6.5 ppm in liver and 303-7100 ppm in kidney). Treatment with MEL and CYA at 1, 3 or 10 mg/kg bw/day for 7 days did not result in residues greater than the level of concern in all tissues tested. Pigs dosed with 33 mg/kg bw/day of MEL + CYA for 7 days contained residues above the level of concern only in kidney. Deposition of MEL and CYA depends on the tissue type (muscles, liver and kidney), dosage and whether the triazines are given alone or in combination. © 2015 .


Hadler J.L.,Yale University | Clogher P.,Yale University | Hurd S.,Yale University | Mandour M.,State Laboratory | And 2 more authors.
Clinical Infectious Diseases | Year: 2011

Background. The epidemiology over time of non-O157 Shiga toxin-producing Escherichia coli (STEC) is unknown. Since 1999, increasing numbers of laboratories in Connecticut have been testing for ST rather than culturing for O157, enabling identification of non-O157 STEC. Methods. Beginning in 2000, Connecticut laboratories were required to submit ST-positive broths to the State Laboratory for isolation and typing of STEC. The ratio of non-O157:O157 from laboratories conducting ST testing was used to determine state-level estimates for non-O157 STEC. Patients with STEC were interviewed for exposure factors in the 7 days preceding illness. Incidence trends, clinical features, and epidemiology of non-O157 and O157 STEC infections were compared. Results. From 1 January 2000 through 31 December 2009, ST testing detected 392 (59%) of 663 reported STEC infections; 229 (58%) of the isolates were non-O157. The estimated incidence of STEC infection decreased by 34%. O157 and the top 4 non-O157 serogroups, O111, O103, O26, and O45, were a stable percentage of all STEC isolates over the 10-year period. Bloody diarrhea, hospitalization, and hemolytic uremic syndrome were more common in patients with O157 STEC than in patients with non-O157 STEC infection. Exposure risks of patients with non-O157 STEC infection differed from those of patients with O157 STEC infection primarily in international travel (15.3% vs 2.5%; P <. 01). Non-O157 types differed from each other with respect to several epidemiologic and exposure features. Conclusions. Both O157 and non-O157 STEC infection incidence decreased from 2000 through 2009. Although infection due to O157 is the most common and clinically severe STEC infection, it accounts for a minority of all clinically significant STEC infections. STEC appear to be a diverse group of organisms that have some differences as well as many epidemiologic and exposure features in common. © 2011 The Author Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved.


PubMed | U.S. Food and Drug Administration, Washington State Public Health Laboratory, State Laboratory and New Hampshire State Public Health Laboratory
Type: | Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association | Year: 2015

Four LC-MS/MS methods were developed to quantify melamine (MEL) and cyanuric acid (CYA) in various pig tissues at or above the level of concern (2.5mg/kg). Pigs treated with 200mg/kg bw/day CYA daily for 7 days did not accumulate significant residue concentrations in muscle, liver or kidney. Pigs treated with 200mg/kg bw MEL daily for 7 or 28 days had MEL residues in muscles (3-13ppm), liver (2.8-14.1ppm) and kidney (9.4-27.2ppm). Treatment with MEL and CYA at 100mg/kg bw of each triazine daily for 7 days resulted in MEL (26-59ppm in muscle, 30-49ppm in liver and 367-6300ppm in kidney) and CYA (1.8-5.8ppm in muscle, 2.6-6.5ppm in liver and 303-7100ppm in kidney). Treatment with MEL and CYA at 1, 3 or 10mg/kg bw/day for 7 days did not result in residues greater than the level of concern in all tissues tested. Pigs dosed with 33mg/kg bw/day of MEL+CYA for 7 days contained residues above the level of concern only in kidney. Deposition of MEL and CYA depends on the tissue type (muscles, liver and kidney), dosage and whether the triazines are given alone or in combination.


PubMed | Friedrich Loeffler Institute, Leibniz Center for Agricultural Landscape Research, State Laboratory, Bernhard Nocht Institute for Tropical Medicine and Zoological Garden Berlin AG
Type: | Journal: Veterinary microbiology | Year: 2016

Usutu virus (USUV) is an arbovirus within the genus flavivirus, which was first introduced to Southern Europe approximately twenty years ago causing epizootics among wild and captive birds. In Germany USUV was initially discovered in wild birds, mainly Common blackbirds (Turdus merula), in the Upper Rhine valley in southwest of the country in 2011 and has not spread much northwards since. Phylogenetic analyses revealed that the still ongoing USUV epidemic is caused by two different USUV strains, USUV-Germany belonging to the USUV Europe 3 lineage and USUV-Bonn belonging to the USUV Africa 3 lineage. The two strains were introduced independently. In August 2015 a new USUV strain, named USUV-Berlin, was isolated in Vero cells from two carcasses of juvenile Great grey owls (Strix nebulosa) kept in the Zoological Garden Berlin, which had suffered from a hyperacute fatal systemic infection. Both owls carried high USUV genome loads. Full-length USUV genomes sequences were determined and phylogenetic analysis demonstrated a close relationship with a Spanish mosquito-derived sequence from 2006. Immunohistochemical antigen detection in organ samples of the owls showed the typical USUV infection patterns. According to the phylogenetic analysis, USUV-Berlin belongs to the Africa 2 lineage, and can thus be distinguished from the other strains circulating in Germany. Repeated findings of different USUV strains suggest more frequent introductions into Central Europe and a higher mobility of this virus than assumed to date.


Malone E.M.,State Laboratory | Malone E.M.,Queen's University of Belfast | Elliott C.T.,Queen's University of Belfast | Kennedy D.G.,Agri Food and Biosciences Institute of Northern Ireland | Regan L.,State Laboratory
Journal of AOAC International | Year: 2010

An LC/MS/MS method was developed and validated for the simultaneous identification, confirmation, and quantification of 12 glucocorticoids in bovine milk. The method was validated in accordance with the criteria defined in Commission Decision 2002/657/EC. The developed method can detect and confirm the presence of dexamethasone, betamethasone, prednisolone, flumethasone, 6α-methylprednisolone, fluorometholone, triamcinolone acetonide, prednisone, cortisone, hydrocortisone, clobetasol propionate, and clobetasol butyrate in bovine milk. Milk samples are extracted with acetonitrile; sodium chloride is subsequently added to aid partition of the milk and acetonitrile mixture. The acetonitrile extract is then subjected to liquid-liquid purification by the addition of hexane. The purified extract is evaporated to dryness and reconstituted in a water-acetonitrile mixture, and determination is carried out by LC/MS/MS. The method permits analysis of up to 30 samples in 1 day.


Power J.D.,St James's Hospital | Kavanagh P.,St James's Hospital | Mclaughlin G.,St James's Hospital | O'Brien J.,Trinity College Dublin | And 5 more authors.
Drug Testing and Analysis | Year: 2015

4-Methylmethcathinone (2-methylamino-1-(4-methylphenyl)propan-1-one, mephedrone) is a psychoactive substance that has been associated with recreational use worldwide. Analytical data related to mephedrone are abundantly available but the characterization of by-products obtained during organic synthesis remains to be explored. This study presents the identification of a 1,2,3,5-tetramethyl-4-(4-methylphenyl)-1H-imidazol-3-ium salt (TMMPI), which was formed during the synthesis of mephedrone. When diethyl ether was added to the crude reaction product, solid material precipitated from the solution. Analytical characterization of TMMPI employed a range of analytical techniques including chromatographic analysis in combination with various mass spectrometric detection methods, nuclear magnetic resonance spectroscopy, and crystal structure analysis. Additional confirmation was obtained from organic synthesis of the imidazolium by-product. When TMMPI was subjected to analysis by gas chromatography-mass spectrometry (GC-MS), isomerization and degradation into two distinct compounds were observed, which pointed towards thermal instability under GC conditions. A liquid chromatography-mass spectrometry (LC-MS) based investigation into a micro-scale synthesis of mephedrone and three additional analogues revealed that the corresponding TMMPI analogue was formed. Interestingly, storage of mephedrone freebase in a number of organic solvents also gave rise to TMMPI and it appeared that its formation during storage was significantly reduced in the absence of air. The present study aimed to support clandestine forensic investigations by employing analytical strategies that are applicable to manufacturing sites. The imidazolium salts will most likely be found amongst the waste products of any clandestine lab site under investigation rather than with the desired product. © 2015 John Wiley & Sons, Ltd.


PubMed | Liverpool John Moores University, State Laboratory, St James's Hospital and Trinity College Dublin
Type: Journal Article | Journal: Drug testing and analysis | Year: 2015

4-Methylmethcathinone (2-methylamino-1-(4-methylphenyl)propan-1-one, mephedrone) is a psychoactive substance that has been associated with recreational use worldwide. Analytical data related to mephedrone are abundantly available but the characterization of by-products obtained during organic synthesis remains to be explored. This study presents the identification of a 1,2,3,5-tetramethyl-4-(4-methylphenyl)-1H-imidazol-3-ium salt (TMMPI), which was formed during the synthesis of mephedrone. When diethyl ether was added to the crude reaction product, solid material precipitated from the solution. Analytical characterization of TMMPI employed a range of analytical techniques including chromatographic analysis in combination with various mass spectrometric detection methods, nuclear magnetic resonance spectroscopy, and crystal structure analysis. Additional confirmation was obtained from organic synthesis of the imidazolium by-product. When TMMPI was subjected to analysis by gas chromatography-mass spectrometry (GC-MS), isomerization and degradation into two distinct compounds were observed, which pointed towards thermal instability under GC conditions. A liquid chromatography-mass spectrometry (LC-MS) based investigation into a micro-scale synthesis of mephedrone and three additional analogues revealed that the corresponding TMMPI analogue was formed. Interestingly, storage of mephedrone freebase in a number of organic solvents also gave rise to TMMPI and it appeared that its formation during storage was significantly reduced in the absence of air. The present study aimed to support clandestine forensic investigations by employing analytical strategies that are applicable to manufacturing sites. The imidazolium salts will most likely be found amongst the waste products of any clandestine lab site under investigation rather than with the desired product.


McDonald M.,Central Meat Control Laboratory | Malone E.,State Laboratory | McBrtde J.,State Laboratory
Journal of AOAC International | Year: 2010

A novel and rapid method was developed and validated for the confirmation of endogenous and synthetic hormones in animal serum using LC/MS/MS. Detection of 17 β-estradiol and β-testosterone below the respective European Union-recommended levels of 0.1 and 0.5 μg/L was achieved, as was a required performance level of 0.1 μg/L for 17 a-estradiol and 0.5 μg/L for 17 a-testosterone, medroxyprogesterone17-acetate, and progesterone. The method was established with dilution of serum followed by ionexchange SPE, LC separation and MS detection with electrospray ionization, selected reaction monitoring, and positive/negative switching. Two characteristic transitions were monitored for each analyte. The method was applied to bovine, ovine, porcine, equine, and avian samples and validated according to European Commission Decision 2002/657/EC and accepted for ISO/IEC 17025:2005 accreditation. An extended calibration curve allows naturally occurring levels of endogenous hormones to be quantified. Recoveries ranged from 97.3% for 17 a-testosterone to 102.0% for 17 a-estradiol. The decision limit CCa ranged from 0.02 ug/L for 17 a- and β-estradiol to 0.12 μg/L for progesterone. Detection capability CCβ ranged from 0.03 μg/L for 17 a-estradiol to 0.20 μg/L for progesterone.


A rapid method has been developed to analyse morphine, codeine, 6-monoacetylmorphine, cocaine, benzoylecgonine, dihydrocodeine, cocaethylene, 3,4-methylenedioxyamphetamine, ketamine, 3,4-methylenedioxymethamphetamine, pseudoephedrine, lignocaine, benzylpiperazine, methamphetamine, amphetamine, methadone, phenethylamine and levamisole in human blood. Blood samples were cleaned up using mixed mode solid phase extraction using Evolute™ CX solid phase extraction cartridges and the sample aliquots were analysed by hybrid triple quadrupole linear ion trap (QTRAP) mass spectrometry with a runtime of 12.5. min. Multiple reaction monitoring (MRM) as survey scan and an enhanced product ion (EPI) scan as dependent scan were performed in an information-dependent acquisition (IDA) experiment. Finally, drug identification and confirmation was carried out by library search with a developed in-house MS/MS library based on EPI spectra at a collision energy spread of 35 ± 15 in positive mode and MRM ratios. The method was validated in blood, according to the criteria defined in Commission Decision 2002/657/EC. At least two MRM transitions for each substance were monitored in addition to EPI spectra. Deuterated analogues of analytes were used as internal standards for quantitation where possible. The method proved to be simple and time efficient and was implemented as an analytical strategy for the illicit drug monitoring of opioids, cocaines, amphetamines and adulterants in forensic cases of crime offenders, abusers or victims in the Republic of Ireland. © 2010 Elsevier B.V.

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