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Antonio Tagarelli - One of the best experts on this subject based on the ideXlab platform.
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development of a simple and rapid solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry method for the analysis of dopamine serotonin and norepinephrine in human urine
Analytica Chimica Acta, 2014Co-Authors: Attilio Naccarato, Giovanni Sindona, Emanuela Gionfriddo, Antonio TagarelliAbstract:Abstract The work aims at developing a simple and rapid method for the quantification of dopamine (DA), serotonin (5-HT) and norepinephrine (NE) in human urine. The urinary levels of these biogenic amines can be correlated with several pathological conditions concerning heart disease, stress, neurological disorders and cancerous tumors. The proposed analytical approach is based on the use of solid phase microextraction (SPME) combined with gas chromatography–triple Quadrupole Mass Spectrometry (GC–QqQ-MS) after a fast derivatization of both aliphatic amino and phenolic moieties by propyl chloroformate. The variables influencing the derivatization reaction were reliably optimized by the multivariate approach of “Experimental design”. The optimal conditions were obtained by performing derivatization with 100 μL of propyl chloroformate and 100 μL of pyridine. The extraction ability of five commercially available SPME fibers was evaluated in univariate mode and the best results were obtained using the polyacrylate fiber. The variables affecting the efficiency of SPME analysis were again optimized by the multivariate approach of “Experimental design” and, in particular, a central composite design (CCD) was applied. The optimal values were extraction in 45 min at room temperature, desorption temperature at 300 °C, no addition of NaCl. Assay of derivatized analytes was performed by using a gas chromatography–triple Quadrupole Mass Spectrometry (GC–QqQ-MS) system in selected reaction monitoring (SRM) acquisition. An evaluation of all analytical parameters demonstrates that the developed method provides satisfactory results. Indeed, very good linearities were achieved in the tested calibration range with correlation coefficient values of 0.9995, 0.9999 and 0.9997 for DA, 5-HT and NE, respectively. Accuracies and RSDs calculated for between-run and tested at concentrations of 30, 200, and 800 μg L−1 were in the range from 92.8% to 103.0%, and from 0.67 to 4.5%, respectively. Finally, the LOD values obtained can be considered very good (0.587, 0.381 and 1.23 μg L−1 for DA, 5-HT and NE, respectively).
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a reliable and simple method for the assay of neuroendocrine tumor markers in human urine by solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry
Analytica Chimica Acta, 2013Co-Authors: Marcello Monteleone, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:Homovanillic acid (HVA), vanylmandelic acid (VMA), and 5-hydroxyindoleacetic acid (5-HIAA) are the metabolites of some catecholamines such as epinephrine, nor-epinephrine, dopamine and serotonin and their quantification is used in the diagnosis and management of patients with neurocrine tumors. A novel approach in the assay of these biomarkers in human urine samples by solid phase microextraction (SPME) combined with gas chromatography-triple Quadrupole Mass Spectrometry (GC-QqQ-MS) is presented. A preliminary derivatization with ethyl chloroformate/ethanol was used and the corresponding derivatives were then extracted by SPME in immersion mode. The performance of five SPME fibers and three chloroformates were evaluated in univariate mode and the best results were obtained using the polyacrylate fiber and ethyl chloroformate. The variables affecting the efficiency of SPME analysis were optimized by the multivariate approach of "Experimental design" and, in particular, a central composite design (CCD) was applied. The optimum working conditions in terms of response values were achieved by performing analysis at room temperature with addition of NaCl (9.5%) and with an extraction time of 25.8 min. Identification and quantification of analytes were carried out by using a gas chromatography-triple Quadrupole Mass Spectrometry (GC-QqQ MS) system in multiple reaction monitoring (MRM) acquisition. An evaluation of all analytical parameters shows that the proposed method provides satisfactory results. Very good linearities were, in fact, achieved in the tested calibration ranges with correlation coefficient values >0.99 for all the analytes and accuracies and RSDs calculated for between-run and tested at concentrations of 1, 10, and 80 mg L(-1) were ranging from 91.3% to 106.6%, and from 0.5 to 8.9%, respectively. Moreover, the LOD values obtained can be considered very satisfactory (1.3, 0.046 and 24.3 μg L(-1) for HVA, VMA and 5-HIAA, respectively). The developed protocol represents, therefore, a simple, rapid and selective tool for assaying these acidic biomarkers in urine samples for neuroendocrine cancer diagnosis.
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a reliable solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry method for the assay of selenomethionine and selenomethylselenocysteine in aqueous extracts difference between selenized and not enriched selenium potatoes
Analytica Chimica Acta, 2012Co-Authors: Emanuela Gionfriddo, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:A new analytical approach is exploited in the assay of selenium speciation in selenized and not selenium enriched potatoes based on the widely available solid-phase microextraction (SPME) coupled to-GC-triple Quadrupole Mass Spectrometry (SPME-GC-QqQ MS) method. The assay of selenomethionine (SeMet) and selenomethylselenocysteine (SeMeSeCys) in potatoes here reported provides clues to the effectiveness of SPME technique combined with gas chromatography-tandem Mass Spectrometry, which could be of general use. For the exploitation of the GC method, the selected analytes were converted into their N(O,S)-alkoxycarbonyl alkyl esters derivatives by direct treatment with alkyl chloroformate in aqueous extracts. The performance of five SPME fibers and three chloroformates were tested in univariate mode and the best results were obtained using the divinylbenzene/carboxen/polydimethylsiloxane fiber and propylchloroformate. The variables affecting the efficiency of SPME analysis were optimized by the multivariate approach of design of experiment (DoE) and, in particular, a central composite design (CCD) was applied. Tandem Mass Spectrometry in selected reaction monitoring (SRM) has allowed the elimination of matrix interferences, providing reconstructed chromatograms with well-resolved peaks and the achievement of very satisfactory detection and quantification limits. Both precision and recovery of the proposed protocol tested at concentration of 8 and 40 μg kg(-1) (dry matter), offered values ranging from 82.3 to 116.3% and from 8.5 to 13.1% for recovery and precision, respectively. The application of the method to commercial samples of selenized and not selenium enriched potatoes proved that the Se fertilization increases significantly the concentration of these bioavailable selenoamino acids.
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sarcosine as a marker in prostate cancer progression a rapid and simple method for its quantification in human urine by solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry
Analytical and Bioanalytical Chemistry, 2011Co-Authors: Brunella Cavaliere, Barbara Macchione, Marcello Monteleone, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:Sarcosine is an amino acid derivative of N-methylglycine and is involved in the amino acid metabolism and methylation processes that are enriched during prostate cancer progression. It could also serve as a new target to be measured during therapeutic interventions and help in the identification of aggressive tumors for radical treatment. In this study, we present a new urine test that can help early diagnosis of prostate cancer. The method for the quantification of sarcosine in urine consists of a solid-phase microextraction (SPME) step followed by gas chromatography–triple Quadrupole Mass Spectrometry analysis. We used a preliminary derivatization step with ethyl chloroformate/ethanol and the corresponding ester was then extracted by SPME in immersion mode. Several fibers were evaluated and the optimization of the parameters affecting the SPME process was carried out using an experimental design. The optimal values were 20 min extraction time, 10% NaCl, and 270°C using a divinylbenzene/Carboxen/polydimethylsiloxane fiber. The triple Quadrupole analyzer acquired data in selected reaction monitoring mode, allowing us to obtain reconstructed chromatograms with well-defined chromatographic peaks. The accuracy and precision of this method were evaluated at concentrations of 70, 250, and 800 ng/ml and were found to be acceptable. Very satisfactory values (0.10 and 0.16 ng/ml, respectively) were also achieved for the limit of detection and the limit of quantification. The proposed protocol represents a rapid, simple, selective, and sensitive tool to quantify sarcosine in urine samples for prostate cancer diagnosis and for a screening test.
Attilio Naccarato - One of the best experts on this subject based on the ideXlab platform.
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development of a simple and rapid solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry method for the analysis of dopamine serotonin and norepinephrine in human urine
Analytica Chimica Acta, 2014Co-Authors: Attilio Naccarato, Giovanni Sindona, Emanuela Gionfriddo, Antonio TagarelliAbstract:Abstract The work aims at developing a simple and rapid method for the quantification of dopamine (DA), serotonin (5-HT) and norepinephrine (NE) in human urine. The urinary levels of these biogenic amines can be correlated with several pathological conditions concerning heart disease, stress, neurological disorders and cancerous tumors. The proposed analytical approach is based on the use of solid phase microextraction (SPME) combined with gas chromatography–triple Quadrupole Mass Spectrometry (GC–QqQ-MS) after a fast derivatization of both aliphatic amino and phenolic moieties by propyl chloroformate. The variables influencing the derivatization reaction were reliably optimized by the multivariate approach of “Experimental design”. The optimal conditions were obtained by performing derivatization with 100 μL of propyl chloroformate and 100 μL of pyridine. The extraction ability of five commercially available SPME fibers was evaluated in univariate mode and the best results were obtained using the polyacrylate fiber. The variables affecting the efficiency of SPME analysis were again optimized by the multivariate approach of “Experimental design” and, in particular, a central composite design (CCD) was applied. The optimal values were extraction in 45 min at room temperature, desorption temperature at 300 °C, no addition of NaCl. Assay of derivatized analytes was performed by using a gas chromatography–triple Quadrupole Mass Spectrometry (GC–QqQ-MS) system in selected reaction monitoring (SRM) acquisition. An evaluation of all analytical parameters demonstrates that the developed method provides satisfactory results. Indeed, very good linearities were achieved in the tested calibration range with correlation coefficient values of 0.9995, 0.9999 and 0.9997 for DA, 5-HT and NE, respectively. Accuracies and RSDs calculated for between-run and tested at concentrations of 30, 200, and 800 μg L−1 were in the range from 92.8% to 103.0%, and from 0.67 to 4.5%, respectively. Finally, the LOD values obtained can be considered very good (0.587, 0.381 and 1.23 μg L−1 for DA, 5-HT and NE, respectively).
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a reliable and simple method for the assay of neuroendocrine tumor markers in human urine by solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry
Analytica Chimica Acta, 2013Co-Authors: Marcello Monteleone, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:Homovanillic acid (HVA), vanylmandelic acid (VMA), and 5-hydroxyindoleacetic acid (5-HIAA) are the metabolites of some catecholamines such as epinephrine, nor-epinephrine, dopamine and serotonin and their quantification is used in the diagnosis and management of patients with neurocrine tumors. A novel approach in the assay of these biomarkers in human urine samples by solid phase microextraction (SPME) combined with gas chromatography-triple Quadrupole Mass Spectrometry (GC-QqQ-MS) is presented. A preliminary derivatization with ethyl chloroformate/ethanol was used and the corresponding derivatives were then extracted by SPME in immersion mode. The performance of five SPME fibers and three chloroformates were evaluated in univariate mode and the best results were obtained using the polyacrylate fiber and ethyl chloroformate. The variables affecting the efficiency of SPME analysis were optimized by the multivariate approach of "Experimental design" and, in particular, a central composite design (CCD) was applied. The optimum working conditions in terms of response values were achieved by performing analysis at room temperature with addition of NaCl (9.5%) and with an extraction time of 25.8 min. Identification and quantification of analytes were carried out by using a gas chromatography-triple Quadrupole Mass Spectrometry (GC-QqQ MS) system in multiple reaction monitoring (MRM) acquisition. An evaluation of all analytical parameters shows that the proposed method provides satisfactory results. Very good linearities were, in fact, achieved in the tested calibration ranges with correlation coefficient values >0.99 for all the analytes and accuracies and RSDs calculated for between-run and tested at concentrations of 1, 10, and 80 mg L(-1) were ranging from 91.3% to 106.6%, and from 0.5 to 8.9%, respectively. Moreover, the LOD values obtained can be considered very satisfactory (1.3, 0.046 and 24.3 μg L(-1) for HVA, VMA and 5-HIAA, respectively). The developed protocol represents, therefore, a simple, rapid and selective tool for assaying these acidic biomarkers in urine samples for neuroendocrine cancer diagnosis.
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a reliable solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry method for the assay of selenomethionine and selenomethylselenocysteine in aqueous extracts difference between selenized and not enriched selenium potatoes
Analytica Chimica Acta, 2012Co-Authors: Emanuela Gionfriddo, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:A new analytical approach is exploited in the assay of selenium speciation in selenized and not selenium enriched potatoes based on the widely available solid-phase microextraction (SPME) coupled to-GC-triple Quadrupole Mass Spectrometry (SPME-GC-QqQ MS) method. The assay of selenomethionine (SeMet) and selenomethylselenocysteine (SeMeSeCys) in potatoes here reported provides clues to the effectiveness of SPME technique combined with gas chromatography-tandem Mass Spectrometry, which could be of general use. For the exploitation of the GC method, the selected analytes were converted into their N(O,S)-alkoxycarbonyl alkyl esters derivatives by direct treatment with alkyl chloroformate in aqueous extracts. The performance of five SPME fibers and three chloroformates were tested in univariate mode and the best results were obtained using the divinylbenzene/carboxen/polydimethylsiloxane fiber and propylchloroformate. The variables affecting the efficiency of SPME analysis were optimized by the multivariate approach of design of experiment (DoE) and, in particular, a central composite design (CCD) was applied. Tandem Mass Spectrometry in selected reaction monitoring (SRM) has allowed the elimination of matrix interferences, providing reconstructed chromatograms with well-resolved peaks and the achievement of very satisfactory detection and quantification limits. Both precision and recovery of the proposed protocol tested at concentration of 8 and 40 μg kg(-1) (dry matter), offered values ranging from 82.3 to 116.3% and from 8.5 to 13.1% for recovery and precision, respectively. The application of the method to commercial samples of selenized and not selenium enriched potatoes proved that the Se fertilization increases significantly the concentration of these bioavailable selenoamino acids.
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sarcosine as a marker in prostate cancer progression a rapid and simple method for its quantification in human urine by solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry
Analytical and Bioanalytical Chemistry, 2011Co-Authors: Brunella Cavaliere, Barbara Macchione, Marcello Monteleone, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:Sarcosine is an amino acid derivative of N-methylglycine and is involved in the amino acid metabolism and methylation processes that are enriched during prostate cancer progression. It could also serve as a new target to be measured during therapeutic interventions and help in the identification of aggressive tumors for radical treatment. In this study, we present a new urine test that can help early diagnosis of prostate cancer. The method for the quantification of sarcosine in urine consists of a solid-phase microextraction (SPME) step followed by gas chromatography–triple Quadrupole Mass Spectrometry analysis. We used a preliminary derivatization step with ethyl chloroformate/ethanol and the corresponding ester was then extracted by SPME in immersion mode. Several fibers were evaluated and the optimization of the parameters affecting the SPME process was carried out using an experimental design. The optimal values were 20 min extraction time, 10% NaCl, and 270°C using a divinylbenzene/Carboxen/polydimethylsiloxane fiber. The triple Quadrupole analyzer acquired data in selected reaction monitoring mode, allowing us to obtain reconstructed chromatograms with well-defined chromatographic peaks. The accuracy and precision of this method were evaluated at concentrations of 70, 250, and 800 ng/ml and were found to be acceptable. Very satisfactory values (0.10 and 0.16 ng/ml, respectively) were also achieved for the limit of detection and the limit of quantification. The proposed protocol represents a rapid, simple, selective, and sensitive tool to quantify sarcosine in urine samples for prostate cancer diagnosis and for a screening test.
S Walorczyk - One of the best experts on this subject based on the ideXlab platform.
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determination of pesticide residues in samples of green minor crops by gas chromatography and ultra performance liquid chromatography coupled to tandem Quadrupole Mass Spectrometry
Talanta, 2015Co-Authors: S Walorczyk, D Drozdzynski, Roman KierzekAbstract:Abstract A method was developed for pesticide analysis in samples of high chlorophyll content belonging to the group of minor crops. A new type of sorbent, known as ChloroFiltr, was employed for dispersive-solid phase extraction cleanup (dispersive-SPE) to reduce the unwanted matrix background prior to concurrent analysis by gas chromatography and ultra-performance liquid chromatography coupled to tandem Quadrupole Mass Spectrometry (GC–MS/MS and UPLC–MS/MS). Validation experiments were carried out on green, unripe plants of lupin, white mustard and sorghum. The overall recoveries at the three spiking levels of 0.01, 0.05 and 0.5 mg kg−1 fell in the range between 68 and 120% (98% on average) and 72–104% (93% on average) with relative standard deviation (RSD) values between 2 and 19% (7% on average) and 3–16% (6% on average) by GC–MS/MS and UPLC–MS/MS technique, respectively. Because of strong enhancement or suppression matrix effects (absolute values >20%) which were exhibited by about 80% of the pesticide and matrix combinations, acceptably accurate quantification was achieved by using matrix-matched standards. Up to now, the proposed method has been successfully used to study the dissipation patterns of pesticides after application on lupin, white mustard, soya bean, sunflower and field bean in experimental plot trials conducted in Poland.
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development and validation of a multi residue method for the determination of pesticides in honeybees using acetonitrile based extraction and gas chromatography tandem Quadrupole Mass Spectrometry
Journal of Chromatography A, 2009Co-Authors: S Walorczyk, B GnusowskiAbstract:An optimized analytical method employing gas chromatography-tandem Quadrupole Mass Spectrometry (GC-MS/MS) has been developed for the simultaneous screening of roughly 150 pesticides in honeybees suspected of poisoning by pesticides during field spraying. In this work, a sample preparation approach based on acetonitrile extraction followed by dispersive solid-phase extraction (d-SPE) cleanup was implemented and validated for pesticides in honeybees for the first time. The procedure involved homogenization of a 2g sample (23 insects on average) with acetonitrile-water mixture followed by salting out with citrate buffer, magnesium sulphate and sodium chloride. An amount of matrix constituents with limited solubility in acetonitrile was reduced in the extract by precipitation at low-temperature (freezing-out cleanup). Hereafter, d-SPE cleanup was carried out using primary secondary amine (PSA), octadecyl (C18) and graphitized carbon black (GCB). This combination of cleanup steps ensured efficient extract purification. Linearity of the calibration curves was studied using matrix-matched standards in the concentration range between 4 and 500 ng mL(-1) (equivalent to 10 and 1250 ng g(-1)), and coefficients of determination (R(2)) were > or =0.99 for approximately 90% of the targeted compounds. The recovery data were obtained by spiking honeybees samples free of pesticides at three concentration levels of 10, 50, and 500 ng g(-1) (approximately 0.9, 4.3, 43.5 ng per bee). At these spiking levels 47, 77 and 92% of the targeted compounds were recovered, respectively. Generally the recoveries were in the range between 70 and 120% with precision values, expressed as relative standard deviation (RSD) < or =20%. The expanded uncertainty was estimated following a "top down" empirical model as being 28% on average (coverage factor k=2, confidence level 95%). Preliminary results from practical application to analysis of real samples are presented. A total of 25 samples of honeybees from suspected pesticides poisoning incidents were analyzed, in which 10 different pesticides were determined.
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application of gas chromatography tandem Quadrupole Mass Spectrometry to the multi residue analysis of pesticides in green leafy vegetables
Rapid Communications in Mass Spectrometry, 2008Co-Authors: S WalorczykAbstract:A new, sensitive and specific method has been developed for the simultaneous determination of 129 pesticides in lettuce and other green leafy vegetables. The samples were extracted with acetonitrile and co-extractives such as fatty acids and pigments were removed using dispersive solid-phase extraction (dispersive-SPE) with primary secondary amine (PSA) and graphitized carbon black (GCB). All pesticides were analyzed in a single injection gas chromatography/tandem Quadrupole Mass Spectrometry (GC/MS/MS) acquisition method. Two multiple reaction monitoring (MRM) transitions of precursor ions fragmenting into product ions were recorded for the targeted pesticides, thus fulfilling the EU identification points system criteria for the identification of contaminants (2002/657/EC). Calibration curves were determined using matrix-matched standards, and exhibited excellent linearity at two orders of magnitude from 0.005 to 0.5 mg/kg for almost all the pesticides studied (R2 ≥ 0.99). The analytical performance was demonstrated by the analysis of lettuce samples spiked at five concentration levels ranging from 0.005 to 0.5 mg/kg for each pesticide. The recovery and repeatability results satisfied SANCO/2007/3131 criteria (i.e. average recoveries were in the range 70–120% with RSDs ≤20%) for 114 of the 129 pesticides at the 0.005 mg/kg spiking level, and for almost all pesticides at the higher spiking levels. The methodology was applied successfully to identify and quantify pesticide residues in leafy vegetable samples such as lettuce, cabbage and leek. Copyright © 2008 John Wiley & Sons, Ltd.
Brendan J Keely - One of the best experts on this subject based on the ideXlab platform.
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comparison of ultra performance liquid chromatography and high performance liquid chromatography for the determination of priority pesticides in baby foods by tandem Quadrupole Mass Spectrometry
Journal of Chromatography A, 2006Co-Authors: Richard J Fussell, Cristiana C Leandro, Peter Hancock, Brendan J KeelyAbstract:Abstract Determination of 16 priority pesticides and transformation products specified in the EU Baby Food Directive 2003/13/EC has been compared using high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC) coupled to tandem Quadrupole Mass Spectrometry (MS/MS). Prior to analysis, co-extractives were removed from acetonitrile extracts using dispersive solid-phase extraction (SPE) with primary secondary amine (50 mg). Extracts spiked with pesticides at 1 μg kg−1 yielded average recoveries in the range 85–119%, with relative standard deviations less than 17%. The HPLC–MS/MS and UPLC–MS/MS multi-residue methods developed are simple, rapid and suitable for the quantification and confirmation of the 16 priority pesticides in fruit-, potato- and cereal-based baby food at 1 μg kg−1. The major advantages of UPLC, using 1.7 μm particles, over HPLC are the speed of analysis, the narrower peaks (giving increased signal-to-noise ratio) and improved confirmation for the targeted pesticides in the analyses of baby foods.
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evaluation of gas chromatography tandem Quadrupole Mass Spectrometry for the determination of organochlorine pesticides in fats and oils
Journal of Chromatography A, 2005Co-Authors: Katan Patel, Richard J Fussell, Mike T Hetmanski, David M Goodall, Brendan J KeelyAbstract:A gas chromatography-tandem Quadrupole Mass Spectrometry multi-residue method for the analysis of 19 organochlorine pesticides in fats and oils has been developed. Gel permeation chromatography was employed to remove lipid material prior to GC-MS/MS analysis. Average recoveries of the pesticides spiked at 10 and 50 microg kg(-1) into fish oil, pork fat, olive oil and hydrogenated vegetable oil were typically in the range 70-110% with relative standard deviations generally less than 10%. Calculated limits of detection are between 0.1 and 2.0 microg kg(-1) and results obtained for the analysis of proficiency test materials are in good agreement with assigned values. The higher selectivity of the GC-MS/MS compared to electron capture detection and GC-MS in selective ion monitoring mode allowed unambiguous identification and confirmation of all the target pesticides at low microg kg(-1) levels in fats and oils in a single analysis.
Giovanni Sindona - One of the best experts on this subject based on the ideXlab platform.
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development of a simple and rapid solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry method for the analysis of dopamine serotonin and norepinephrine in human urine
Analytica Chimica Acta, 2014Co-Authors: Attilio Naccarato, Giovanni Sindona, Emanuela Gionfriddo, Antonio TagarelliAbstract:Abstract The work aims at developing a simple and rapid method for the quantification of dopamine (DA), serotonin (5-HT) and norepinephrine (NE) in human urine. The urinary levels of these biogenic amines can be correlated with several pathological conditions concerning heart disease, stress, neurological disorders and cancerous tumors. The proposed analytical approach is based on the use of solid phase microextraction (SPME) combined with gas chromatography–triple Quadrupole Mass Spectrometry (GC–QqQ-MS) after a fast derivatization of both aliphatic amino and phenolic moieties by propyl chloroformate. The variables influencing the derivatization reaction were reliably optimized by the multivariate approach of “Experimental design”. The optimal conditions were obtained by performing derivatization with 100 μL of propyl chloroformate and 100 μL of pyridine. The extraction ability of five commercially available SPME fibers was evaluated in univariate mode and the best results were obtained using the polyacrylate fiber. The variables affecting the efficiency of SPME analysis were again optimized by the multivariate approach of “Experimental design” and, in particular, a central composite design (CCD) was applied. The optimal values were extraction in 45 min at room temperature, desorption temperature at 300 °C, no addition of NaCl. Assay of derivatized analytes was performed by using a gas chromatography–triple Quadrupole Mass Spectrometry (GC–QqQ-MS) system in selected reaction monitoring (SRM) acquisition. An evaluation of all analytical parameters demonstrates that the developed method provides satisfactory results. Indeed, very good linearities were achieved in the tested calibration range with correlation coefficient values of 0.9995, 0.9999 and 0.9997 for DA, 5-HT and NE, respectively. Accuracies and RSDs calculated for between-run and tested at concentrations of 30, 200, and 800 μg L−1 were in the range from 92.8% to 103.0%, and from 0.67 to 4.5%, respectively. Finally, the LOD values obtained can be considered very good (0.587, 0.381 and 1.23 μg L−1 for DA, 5-HT and NE, respectively).
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a reliable and simple method for the assay of neuroendocrine tumor markers in human urine by solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry
Analytica Chimica Acta, 2013Co-Authors: Marcello Monteleone, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:Homovanillic acid (HVA), vanylmandelic acid (VMA), and 5-hydroxyindoleacetic acid (5-HIAA) are the metabolites of some catecholamines such as epinephrine, nor-epinephrine, dopamine and serotonin and their quantification is used in the diagnosis and management of patients with neurocrine tumors. A novel approach in the assay of these biomarkers in human urine samples by solid phase microextraction (SPME) combined with gas chromatography-triple Quadrupole Mass Spectrometry (GC-QqQ-MS) is presented. A preliminary derivatization with ethyl chloroformate/ethanol was used and the corresponding derivatives were then extracted by SPME in immersion mode. The performance of five SPME fibers and three chloroformates were evaluated in univariate mode and the best results were obtained using the polyacrylate fiber and ethyl chloroformate. The variables affecting the efficiency of SPME analysis were optimized by the multivariate approach of "Experimental design" and, in particular, a central composite design (CCD) was applied. The optimum working conditions in terms of response values were achieved by performing analysis at room temperature with addition of NaCl (9.5%) and with an extraction time of 25.8 min. Identification and quantification of analytes were carried out by using a gas chromatography-triple Quadrupole Mass Spectrometry (GC-QqQ MS) system in multiple reaction monitoring (MRM) acquisition. An evaluation of all analytical parameters shows that the proposed method provides satisfactory results. Very good linearities were, in fact, achieved in the tested calibration ranges with correlation coefficient values >0.99 for all the analytes and accuracies and RSDs calculated for between-run and tested at concentrations of 1, 10, and 80 mg L(-1) were ranging from 91.3% to 106.6%, and from 0.5 to 8.9%, respectively. Moreover, the LOD values obtained can be considered very satisfactory (1.3, 0.046 and 24.3 μg L(-1) for HVA, VMA and 5-HIAA, respectively). The developed protocol represents, therefore, a simple, rapid and selective tool for assaying these acidic biomarkers in urine samples for neuroendocrine cancer diagnosis.
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a reliable solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry method for the assay of selenomethionine and selenomethylselenocysteine in aqueous extracts difference between selenized and not enriched selenium potatoes
Analytica Chimica Acta, 2012Co-Authors: Emanuela Gionfriddo, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:A new analytical approach is exploited in the assay of selenium speciation in selenized and not selenium enriched potatoes based on the widely available solid-phase microextraction (SPME) coupled to-GC-triple Quadrupole Mass Spectrometry (SPME-GC-QqQ MS) method. The assay of selenomethionine (SeMet) and selenomethylselenocysteine (SeMeSeCys) in potatoes here reported provides clues to the effectiveness of SPME technique combined with gas chromatography-tandem Mass Spectrometry, which could be of general use. For the exploitation of the GC method, the selected analytes were converted into their N(O,S)-alkoxycarbonyl alkyl esters derivatives by direct treatment with alkyl chloroformate in aqueous extracts. The performance of five SPME fibers and three chloroformates were tested in univariate mode and the best results were obtained using the divinylbenzene/carboxen/polydimethylsiloxane fiber and propylchloroformate. The variables affecting the efficiency of SPME analysis were optimized by the multivariate approach of design of experiment (DoE) and, in particular, a central composite design (CCD) was applied. Tandem Mass Spectrometry in selected reaction monitoring (SRM) has allowed the elimination of matrix interferences, providing reconstructed chromatograms with well-resolved peaks and the achievement of very satisfactory detection and quantification limits. Both precision and recovery of the proposed protocol tested at concentration of 8 and 40 μg kg(-1) (dry matter), offered values ranging from 82.3 to 116.3% and from 8.5 to 13.1% for recovery and precision, respectively. The application of the method to commercial samples of selenized and not selenium enriched potatoes proved that the Se fertilization increases significantly the concentration of these bioavailable selenoamino acids.
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sarcosine as a marker in prostate cancer progression a rapid and simple method for its quantification in human urine by solid phase microextraction gas chromatography triple Quadrupole Mass Spectrometry
Analytical and Bioanalytical Chemistry, 2011Co-Authors: Brunella Cavaliere, Barbara Macchione, Marcello Monteleone, Attilio Naccarato, Giovanni Sindona, Antonio TagarelliAbstract:Sarcosine is an amino acid derivative of N-methylglycine and is involved in the amino acid metabolism and methylation processes that are enriched during prostate cancer progression. It could also serve as a new target to be measured during therapeutic interventions and help in the identification of aggressive tumors for radical treatment. In this study, we present a new urine test that can help early diagnosis of prostate cancer. The method for the quantification of sarcosine in urine consists of a solid-phase microextraction (SPME) step followed by gas chromatography–triple Quadrupole Mass Spectrometry analysis. We used a preliminary derivatization step with ethyl chloroformate/ethanol and the corresponding ester was then extracted by SPME in immersion mode. Several fibers were evaluated and the optimization of the parameters affecting the SPME process was carried out using an experimental design. The optimal values were 20 min extraction time, 10% NaCl, and 270°C using a divinylbenzene/Carboxen/polydimethylsiloxane fiber. The triple Quadrupole analyzer acquired data in selected reaction monitoring mode, allowing us to obtain reconstructed chromatograms with well-defined chromatographic peaks. The accuracy and precision of this method were evaluated at concentrations of 70, 250, and 800 ng/ml and were found to be acceptable. Very satisfactory values (0.10 and 0.16 ng/ml, respectively) were also achieved for the limit of detection and the limit of quantification. The proposed protocol represents a rapid, simple, selective, and sensitive tool to quantify sarcosine in urine samples for prostate cancer diagnosis and for a screening test.
