Monica Mazzarino - Academia.edu (original) (raw)

Papers by Monica Mazzarino

Research paper thumbnail of UHPLC–HRMS Method for the Simultaneous Screening of 235 Drugs in Capillary Blood for Doping Control Purpose: Comparative Evaluation of Volumetric and Non-volumetric Dried Blood Spotting Devices

ACS omega, Aug 29, 2022

We present a quick and simple multi-targeted analytical workflow based on ultra-high-performance ... more We present a quick and simple multi-targeted analytical workflow based on ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry for the screening in dried blood spots and dried plasma spots of a wide variety of drugs with different chemical properties. Seven different microsampling devices were evaluated in view of their application for the detection of the selected target analytes in the framework of doping control analysis. The extraction of the analytes was optimized by assessing the efficacy of protocols based on ultrasonication with aqueous buffers and/or organic solvents of different polarities. Optimal recoveries were obtained by using pure methanol or mixtures of methanol/acetonitrile and methanol/isopropanol, depending on both the device and the target analytes. The method was fully validated according to both ISO17025 and the requirements of the World Anti-Doping Agency: all the analytes were clearly distinguishable from the matrix, with limits of detection in the range of 0.1–3.0 ng mL–1. Stability studies simulating the storage of samples before the analysis and in view of a possible re-analysis showed that most of the analytes were stable for at least 24 h at 50 °C and for at least 3 weeks at 25 and at 4 °C. The real applicability of the method was assessed by analyzing the samples collected after the administration of two model drugs, acetazolamide and deflazacort. The performance of the method was confirmed to be fit for purpose, and data obtained in blood can also be used to complement those available in urine, allowing to refine the knowledge concerning the pharmacokinetic profiles.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of A screening method for the simultaneous detection of glucocorticoids, diuretics, stimulants, anti-oestrogens, beta-adrenergic drugs and anabolic steroids in human urine by LC-ESI-MS/MS

Analytical and Bioanalytical Chemistry, Aug 14, 2008

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Rapid determination of diuretics in human urine by gas chromatography–mass spectrometry following microwave assisted derivatization

Analytica Chimica Acta, 2003

This work presents a complete method for the screening and confirmation analysis of diuretics in ... more This work presents a complete method for the screening and confirmation analysis of diuretics in human urine by gas chromatography–mass spectrometry (GC–MS). The method comprises a pretreatment stage (extraction, preconcentration and derivatization to form ...

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Effect of the systemic versus inhalatory administration of synthetic glucocorticoids on the urinary steroid profile as studied by gas chromatography–mass spectrometry

Analytica Chimica Acta, Feb 1, 2006

This paper presents a gas chromatography–mass spectrometry (GC–MS) study carried out on human uri... more This paper presents a gas chromatography–mass spectrometry (GC–MS) study carried out on human urine to verify whether the administration of glucocorticoids can affect the urinary steroid profile, and especially the levels of endogenous glucocorticoids, androgens and their main metabolites.Betamethasone and beclomethasone, administered either systemically (per os or i.m.) or locally (by inhalation) have been studied. The determination of the urinary

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

Forensic Toxicology, May 27, 2015

A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously ... more A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously detect 19 small peptide hormones prohibited in sport and their main metabolites in urine after solid-phase extraction. Detection was achieved using a triple-quadrupole mass spectrometric detector coupled with an electrospray ionization interface after chromatographic separation with an octadecyl column based on fused-core particle technology. Sample pretreatment was performed by solid-phase extraction. The extraction procedure was optimized by comparison of different sorbents and washing/elution protocols. The best results were obtained using a mixed-mode weak cation exchange sorbent, two washing steps (ultrapurified water and methanol), and elution using 300 mM ammonium formate in 25 % ammonia/methanol (10/90) or 25 % ammonia/10 % formic acid/methanol (8/12/80). The procedure was validated in terms of sensitivity (lower limits of detection: 0.05–2.0 ng/ml depending on the target analyte), specificity, recovery [>60 %, coefficient of variation (CV) <15 % except for TB500 17–23 fragment, AOD9604, and ARA290 for which recovery was <50 %), ion suppression/enhancement (<35 %), robustness, carryover, stability of the target analytes [stable for at least for 2 days (25 °C, 2 weeks (4 °C), 2 months (−20 °C)], and repeatability of retention times (CV <0.1 %) and relative abundances of the selected ion transitions (CV <15 %). The suitability of the method was confirmed by analyzing spiked and excreted urines, the latter collected after intravenous injection of 0.1 mg of GHRP-2.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of A fast liquid chromatographic/mass spectrometric screening method for the simultaneous detection of synthetic glucocorticoids, some stimulants, anti-oestrogen drugs and synthetic anabolic steroids

Rapid Communications in Mass Spectrometry, 2006

A fast liquid chromatographic/mass spectrometric (LC/MS/MS) screening method for the detection, i... more A fast liquid chromatographic/mass spectrometric (LC/MS/MS) screening method for the detection, in urine, of synthetic glucocorticoids, stimulants (formoterol, modafinil and mesocarb), anti-oestrogens (finasteride, exemestane, anastrozole, letrozole and formestane) and synthetic anabolic steroids (stanozolol, gestrinone and tetrahydrogestrinone) is described. All these drugs (and/or their urinary metabolites) can be simultaneously extracted by a single liquid/liquid extraction step, at alkaline pH, after enzymatic hydrolysis with beta-glucuronidase, and assayed in 7 min by LC/MS/MS using electrospray ionization in positive ion mode and multiple reaction monitoring as the acquisition mode. All compounds show good reproducibility of both the retention times (CV% &amp;amp;amp;amp;lt;2%) and the relative abundances (CV% &amp;amp;amp;amp;lt;10%). The limits of detection for the anti-oestrogens, glucocorticoids and steroids are in the range of 1-30 ng/mL, and for the stimulants are in the range of 100-200 ng/mL, thus satisfying the minimum required performance limits of the World Anti-Doping Agency.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Development and validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of phthalates and bisphenol a in serum, urine and follicular fluid

Clinical Mass Spectrometry, Nov 1, 2020

Phthalates and bisphenol A interfere with the synthesis, secretion, transport, binding, metabolis... more Phthalates and bisphenol A interfere with the synthesis, secretion, transport, binding, metabolism, and excretion of endogenous hormones and, for this reason, are classified as endocrine disruptors.We are here presenting an analytical method for the simultaneous detection of six phthalates metabolites and bisphenol A in different biological fluids (urine, serum and follifular fluid) by liquid chromatography coupled to tandem mass spectrometry. The quantification was carried out in negative electrospray ionization mode using selected reaction monitoring as acquisition mode. Different extraction protocols, using either solid phase or liquid/liquid extraction, were comparatively evaluated to optimize the sample preparation procedure. Solid-phase extraction was chosen as it ensured the best recovery and overall sensitivity. The method was successfully validated: recovery varying in the range 71 ± 2%–107 ± 6% depending on the target analyte and the matrix considered, intra-assay and inter-assay precision ≤ 12% for follicular fluid, ≤11% for serum and ≤ 10% for urine and accuracy ≤ 115% for follicular fluid, ≤113% for serum ≤ 115% for urine , linearity with R2 > 0.99, with the exception of MEP (recovery 64 ± 8%, intra-assay precision ≤ 20%, inter-assay precision ≤ 16% for follicular fluid). The actual applicability of the method developed and validated in this study was assessed by the analysis of real samples, including 10 specimens of follicular fluid, serum and urine samples, that showed the presence of phthalates metabolites and Bisphenol A, and confirming that the newly developed method can be applied in the routine clinical laboratory for the identification and quantitation of these endocrine-disrupting chemicals.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Capillary blood as a complementary matrix for doping control purposes. Application to the definition of the individual longitudinal profile of IGF-1

Journal of Pharmaceutical and Biomedical Analysis

Bookmarks Related papers MentionsView impact

Research paper thumbnail of In vitro metabolic profile of mexedrone, a mephedrone analog, studied by high‐ and low‐resolution mass spectrometry

Drug Testing and Analysis, 2021

Mexedrone is a synthetic cathinone structurally related to mephedrone, which belongs to the class... more Mexedrone is a synthetic cathinone structurally related to mephedrone, which belongs to the class of N‐alkyl cathinone derivatives, whose metabolic profile has not been fully clarified yet. This study considers the in vitro phase I metabolism of mexedrone, to pre‐select the most appropriate marker(s) of intake. Mexedrone was incubated in the presence of either human liver microsomes or single recombinant CYP450 isoforms. The metabolic profile was outlined by ultra‐high‐performance liquid chromatography coupled to both high‐ and low‐resolution mass spectrometry. In detail, the phase I metabolic profile of mexedrone was initially defined by a time‐of‐flight analyzer, while the chemical structures of the detected metabolites and the potential presence of minor metabolites were subsequently studied by tandem mass spectrometry, using a triple quadrupole analyzer. The main phase I metabolic reactions were hydroxylation and N‐ and O‐dealkylation. The CYP450 isoforms most involved were CYP2...

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Urinary excretion profile of methiopropamine in mice following intraperitoneal administration: A liquid chromatography–tandem mass spectrometry investigation

Drug Testing and Analysis, 2020

We have considered the urinary excretion profile of methiopropamine (MPA), a thiophene ring‐based... more We have considered the urinary excretion profile of methiopropamine (MPA), a thiophene ring‐based structural analog of methamphetamine with similar stimulant effects, with the aim of selecting the most appropriate marker(s) of intake that may be useful in forensic analysis. For this purpose, in vitro studies were preliminarily performed on human liver microsomes for tracing the phase I metabolic pathways of MPA, preselecting the best candidates as potential target analytes, and designing the optimal experimental strategy. In vivo studies were then conducted on mice, after the intraperitoneal administration of a 10‐mg/kg dose. Urine samples were collected every 3 h in the first 9 h and, subsequently, from 24 to 36 h, and stored at –80°C until further analysis. The measurements were performed using a targeted procedure based on liquid/liquid extraction followed by liquid chromatography–tandem mass spectrometry analysis. Our results show that in the time interval 0–9 h after administra...

Bookmarks Related papers MentionsView impact

Research paper thumbnail of How reliable is dietary supplement labelling?—Experiences from the analysis of ecdysterone supplements

Journal of Pharmaceutical and Biomedical Analysis, 2019

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Urinary and serum hormones profiles after testosterone enanthate administration in male hypogonadism: Concerns on the detection of doping with testosterone in treated hypogonadal athletes

Journal of Endocrinological Investigation, 2009

To describe serum and urinary hormones, androgens metabolites and testosterone/epitestosterone ra... more To describe serum and urinary hormones, androgens metabolites and testosterone/epitestosterone ratio profiles after testosterone administration in male hypogonadal volunteers, and to evaluate their possible usefulness in detecting doping with testosterone in treated hypogonadal athletes. Controlled open label design vs placebo; pharmacokinetic study. Ten male volunteers affected by severe hypogonadism (serum testosterone &amp;amp;amp;amp;amp;amp;amp;amp;lt;2.31 ng/ml). Serum and urinary parameters were evaluated, by radioimmunoassay and gas chromatography-mass spectrometry, before and at different time points for 7/3 weeks after a single administration of testosterone enanthate (250 mg) or placebo, respectively. As partially known, testosterone administration increased, with great individual variability, urinary concentrations of glucuronide testosterone, androsterone, etiocholanolone, 5alpha-androstane- 3alpha,17beta-diol, 5beta-androstane-3alpha,17beta-diol, testosterone/ epitestosterone and testosterone/LH ratios; and decreased epitestosterone and 5alpha-androstane-3beta,17beta-diol/5beta-androstane- 3alpha,17beta-diol ratio. Serum testosterone and dihydrotestosterone increased in all volunteers, and concentrations higher than the upper reference limits were observed in many volunteers until 2 weeks after testosterone administration. Whereas the observed prolonged hyperandrogenism partially limited data interpretation, the report ed characteristics of variation of urinary parameters might be used to suspect testosterone misuse in hypogonadal athletes treated with testosterone enanthate. In this sense, while the actual threshold for tes tos terone/epites tos ter one ratio was confirmed to be of reduced usefulness, we suggest a contemporary evaluation of whole urinary androgen metabolites profile and serum androgens, at specific time points after testosterone enanthate administration. Moreover, an adequate tailoring of treatment, to avoid transitory hyperandrogenism, is highly advisable. Further studies on strategies for detecting doping with testosterone in hypogonadal athletes are warranted.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

Forensic Toxicology, 2015

A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously ... more A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously detect 19 small peptide hormones prohibited in sport and their main metabolites in urine after solid-phase extraction. Detection was achieved using a triple-quadrupole mass spectrometric detector coupled with an electrospray ionization interface after chromatographic separation with an octadecyl column based on fused-core particle technology. Sample pretreatment was performed by solid-phase extraction. The extraction procedure was optimized by comparison of different sorbents and washing/elution protocols. The best results were obtained using a mixed-mode weak cation exchange sorbent, two washing steps (ultrapurified water and methanol), and elution using 300 mM ammonium formate in 25 % ammonia/methanol (10/90) or 25 % ammonia/10 % formic acid/methanol (8/12/80). The procedure was validated in terms of sensitivity (lower limits of detection: 0.05–2.0 ng/ml depending on the target analyte), specificity, recovery [>60 %, coefficient of variation (CV) <15 % except for TB500 17–23 fragment, AOD9604, and ARA290 for which recovery was <50 %), ion suppression/enhancement (<35 %), robustness, carryover, stability of the target analytes [stable for at least for 2 days (25 °C, 2 weeks (4 °C), 2 months (−20 °C)], and repeatability of retention times (CV <0.1 %) and relative abundances of the selected ion transitions (CV <15 %). The suitability of the method was confirmed by analyzing spiked and excreted urines, the latter collected after intravenous injection of 0.1 mg of GHRP-2.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of A screening method for the detection of synthetic glucocorticosteroids in human urine by liquid chromatography–mass spectrometry based on class-characteristic fragmentation pathways

Analytical and Bioanalytical Chemistry, Feb 9, 2008

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Detection of synthetic analogues of insulin-like growth factor 1 in different biological fluids by liquid chromatography quadrupole time-of-flight mass spectrometry: comparison of different immunoaffinity protocols

Analytical and Bioanalytical Chemistry

Bookmarks Related papers MentionsView impact

Research paper thumbnail of UHPLC–HRMS Method for the Simultaneous Screening of 235 Drugs in Capillary Blood for Doping Control Purpose: Comparative Evaluation of Volumetric and Non-volumetric Dried Blood Spotting Devices

ACS Omega

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Urinary excretion profile of prednisolone and prednisone after rectal administration: significance in antidoping analysis

Drug Testing and Analysis

Bookmarks Related papers MentionsView impact

Research paper thumbnail of How safe are our internal procedures? Some preliminary experimental evidence on the problem of “lab-oriented” potential masking agents

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Drug-drug interactions and masking effects in sport doping: influence of miconazole administration on the urinary concentrations of endogenous anabolic steroids

Forensic Toxicology, 2016

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Human hepatoma cell lines on gas foaming templated alginate scaffolds for in vitro drug-drug interaction and metabolism studies

Toxicology in vitro : an international journal published in association with BIBRA, Jan 6, 2015

Liver in vitro systems that allow reliable prediction of major human in vivo metabolic pathways h... more Liver in vitro systems that allow reliable prediction of major human in vivo metabolic pathways have a significant impact in drug screening and drug metabolism research. In the present study, a novel porous scaffold composed of alginate was prepared by employing a gas-in-liquid foaming approach. Galactose residues were introduced on scaffold surfaces to promote cell adhesion and to enhance liver specific functions of the entrapped HepG2/C3A cells. Hepatoma cells in the gal-alginate scaffold showed higher levels of liver specific products (albumin and urea) and were more responsive to specific inducers (e.g. dexamethasone) and inhibitors (e.g. ketoconazole) of the CYP3A4 system than in conventional monolayer culture. HepG2/C3A cells were also more efficient in terms of rapid elimination of testosterone, used as a model substance, at rates comparable to those of in vivo excretion. In addition, an improvement in metabolism of testosterone, in terms of phase II metabolite formation, was...

Bookmarks Related papers MentionsView impact

Research paper thumbnail of UHPLC–HRMS Method for the Simultaneous Screening of 235 Drugs in Capillary Blood for Doping Control Purpose: Comparative Evaluation of Volumetric and Non-volumetric Dried Blood Spotting Devices

ACS omega, Aug 29, 2022

We present a quick and simple multi-targeted analytical workflow based on ultra-high-performance ... more We present a quick and simple multi-targeted analytical workflow based on ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry for the screening in dried blood spots and dried plasma spots of a wide variety of drugs with different chemical properties. Seven different microsampling devices were evaluated in view of their application for the detection of the selected target analytes in the framework of doping control analysis. The extraction of the analytes was optimized by assessing the efficacy of protocols based on ultrasonication with aqueous buffers and/or organic solvents of different polarities. Optimal recoveries were obtained by using pure methanol or mixtures of methanol/acetonitrile and methanol/isopropanol, depending on both the device and the target analytes. The method was fully validated according to both ISO17025 and the requirements of the World Anti-Doping Agency: all the analytes were clearly distinguishable from the matrix, with limits of detection in the range of 0.1–3.0 ng mL–1. Stability studies simulating the storage of samples before the analysis and in view of a possible re-analysis showed that most of the analytes were stable for at least 24 h at 50 °C and for at least 3 weeks at 25 and at 4 °C. The real applicability of the method was assessed by analyzing the samples collected after the administration of two model drugs, acetazolamide and deflazacort. The performance of the method was confirmed to be fit for purpose, and data obtained in blood can also be used to complement those available in urine, allowing to refine the knowledge concerning the pharmacokinetic profiles.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of A screening method for the simultaneous detection of glucocorticoids, diuretics, stimulants, anti-oestrogens, beta-adrenergic drugs and anabolic steroids in human urine by LC-ESI-MS/MS

Analytical and Bioanalytical Chemistry, Aug 14, 2008

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Rapid determination of diuretics in human urine by gas chromatography–mass spectrometry following microwave assisted derivatization

Analytica Chimica Acta, 2003

This work presents a complete method for the screening and confirmation analysis of diuretics in ... more This work presents a complete method for the screening and confirmation analysis of diuretics in human urine by gas chromatography–mass spectrometry (GC–MS). The method comprises a pretreatment stage (extraction, preconcentration and derivatization to form ...

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Effect of the systemic versus inhalatory administration of synthetic glucocorticoids on the urinary steroid profile as studied by gas chromatography–mass spectrometry

Analytica Chimica Acta, Feb 1, 2006

This paper presents a gas chromatography–mass spectrometry (GC–MS) study carried out on human uri... more This paper presents a gas chromatography–mass spectrometry (GC–MS) study carried out on human urine to verify whether the administration of glucocorticoids can affect the urinary steroid profile, and especially the levels of endogenous glucocorticoids, androgens and their main metabolites.Betamethasone and beclomethasone, administered either systemically (per os or i.m.) or locally (by inhalation) have been studied. The determination of the urinary

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

Forensic Toxicology, May 27, 2015

A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously ... more A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously detect 19 small peptide hormones prohibited in sport and their main metabolites in urine after solid-phase extraction. Detection was achieved using a triple-quadrupole mass spectrometric detector coupled with an electrospray ionization interface after chromatographic separation with an octadecyl column based on fused-core particle technology. Sample pretreatment was performed by solid-phase extraction. The extraction procedure was optimized by comparison of different sorbents and washing/elution protocols. The best results were obtained using a mixed-mode weak cation exchange sorbent, two washing steps (ultrapurified water and methanol), and elution using 300 mM ammonium formate in 25 % ammonia/methanol (10/90) or 25 % ammonia/10 % formic acid/methanol (8/12/80). The procedure was validated in terms of sensitivity (lower limits of detection: 0.05–2.0 ng/ml depending on the target analyte), specificity, recovery [>60 %, coefficient of variation (CV) <15 % except for TB500 17–23 fragment, AOD9604, and ARA290 for which recovery was <50 %), ion suppression/enhancement (<35 %), robustness, carryover, stability of the target analytes [stable for at least for 2 days (25 °C, 2 weeks (4 °C), 2 months (−20 °C)], and repeatability of retention times (CV <0.1 %) and relative abundances of the selected ion transitions (CV <15 %). The suitability of the method was confirmed by analyzing spiked and excreted urines, the latter collected after intravenous injection of 0.1 mg of GHRP-2.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of A fast liquid chromatographic/mass spectrometric screening method for the simultaneous detection of synthetic glucocorticoids, some stimulants, anti-oestrogen drugs and synthetic anabolic steroids

Rapid Communications in Mass Spectrometry, 2006

A fast liquid chromatographic/mass spectrometric (LC/MS/MS) screening method for the detection, i... more A fast liquid chromatographic/mass spectrometric (LC/MS/MS) screening method for the detection, in urine, of synthetic glucocorticoids, stimulants (formoterol, modafinil and mesocarb), anti-oestrogens (finasteride, exemestane, anastrozole, letrozole and formestane) and synthetic anabolic steroids (stanozolol, gestrinone and tetrahydrogestrinone) is described. All these drugs (and/or their urinary metabolites) can be simultaneously extracted by a single liquid/liquid extraction step, at alkaline pH, after enzymatic hydrolysis with beta-glucuronidase, and assayed in 7 min by LC/MS/MS using electrospray ionization in positive ion mode and multiple reaction monitoring as the acquisition mode. All compounds show good reproducibility of both the retention times (CV% &amp;amp;amp;amp;lt;2%) and the relative abundances (CV% &amp;amp;amp;amp;lt;10%). The limits of detection for the anti-oestrogens, glucocorticoids and steroids are in the range of 1-30 ng/mL, and for the stimulants are in the range of 100-200 ng/mL, thus satisfying the minimum required performance limits of the World Anti-Doping Agency.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Development and validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of phthalates and bisphenol a in serum, urine and follicular fluid

Clinical Mass Spectrometry, Nov 1, 2020

Phthalates and bisphenol A interfere with the synthesis, secretion, transport, binding, metabolis... more Phthalates and bisphenol A interfere with the synthesis, secretion, transport, binding, metabolism, and excretion of endogenous hormones and, for this reason, are classified as endocrine disruptors.We are here presenting an analytical method for the simultaneous detection of six phthalates metabolites and bisphenol A in different biological fluids (urine, serum and follifular fluid) by liquid chromatography coupled to tandem mass spectrometry. The quantification was carried out in negative electrospray ionization mode using selected reaction monitoring as acquisition mode. Different extraction protocols, using either solid phase or liquid/liquid extraction, were comparatively evaluated to optimize the sample preparation procedure. Solid-phase extraction was chosen as it ensured the best recovery and overall sensitivity. The method was successfully validated: recovery varying in the range 71 ± 2%–107 ± 6% depending on the target analyte and the matrix considered, intra-assay and inter-assay precision ≤ 12% for follicular fluid, ≤11% for serum and ≤ 10% for urine and accuracy ≤ 115% for follicular fluid, ≤113% for serum ≤ 115% for urine , linearity with R2 > 0.99, with the exception of MEP (recovery 64 ± 8%, intra-assay precision ≤ 20%, inter-assay precision ≤ 16% for follicular fluid). The actual applicability of the method developed and validated in this study was assessed by the analysis of real samples, including 10 specimens of follicular fluid, serum and urine samples, that showed the presence of phthalates metabolites and Bisphenol A, and confirming that the newly developed method can be applied in the routine clinical laboratory for the identification and quantitation of these endocrine-disrupting chemicals.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Capillary blood as a complementary matrix for doping control purposes. Application to the definition of the individual longitudinal profile of IGF-1

Journal of Pharmaceutical and Biomedical Analysis

Bookmarks Related papers MentionsView impact

Research paper thumbnail of In vitro metabolic profile of mexedrone, a mephedrone analog, studied by high‐ and low‐resolution mass spectrometry

Drug Testing and Analysis, 2021

Mexedrone is a synthetic cathinone structurally related to mephedrone, which belongs to the class... more Mexedrone is a synthetic cathinone structurally related to mephedrone, which belongs to the class of N‐alkyl cathinone derivatives, whose metabolic profile has not been fully clarified yet. This study considers the in vitro phase I metabolism of mexedrone, to pre‐select the most appropriate marker(s) of intake. Mexedrone was incubated in the presence of either human liver microsomes or single recombinant CYP450 isoforms. The metabolic profile was outlined by ultra‐high‐performance liquid chromatography coupled to both high‐ and low‐resolution mass spectrometry. In detail, the phase I metabolic profile of mexedrone was initially defined by a time‐of‐flight analyzer, while the chemical structures of the detected metabolites and the potential presence of minor metabolites were subsequently studied by tandem mass spectrometry, using a triple quadrupole analyzer. The main phase I metabolic reactions were hydroxylation and N‐ and O‐dealkylation. The CYP450 isoforms most involved were CYP2...

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Urinary excretion profile of methiopropamine in mice following intraperitoneal administration: A liquid chromatography–tandem mass spectrometry investigation

Drug Testing and Analysis, 2020

We have considered the urinary excretion profile of methiopropamine (MPA), a thiophene ring‐based... more We have considered the urinary excretion profile of methiopropamine (MPA), a thiophene ring‐based structural analog of methamphetamine with similar stimulant effects, with the aim of selecting the most appropriate marker(s) of intake that may be useful in forensic analysis. For this purpose, in vitro studies were preliminarily performed on human liver microsomes for tracing the phase I metabolic pathways of MPA, preselecting the best candidates as potential target analytes, and designing the optimal experimental strategy. In vivo studies were then conducted on mice, after the intraperitoneal administration of a 10‐mg/kg dose. Urine samples were collected every 3 h in the first 9 h and, subsequently, from 24 to 36 h, and stored at –80°C until further analysis. The measurements were performed using a targeted procedure based on liquid/liquid extraction followed by liquid chromatography–tandem mass spectrometry analysis. Our results show that in the time interval 0–9 h after administra...

Bookmarks Related papers MentionsView impact

Research paper thumbnail of How reliable is dietary supplement labelling?—Experiences from the analysis of ecdysterone supplements

Journal of Pharmaceutical and Biomedical Analysis, 2019

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Urinary and serum hormones profiles after testosterone enanthate administration in male hypogonadism: Concerns on the detection of doping with testosterone in treated hypogonadal athletes

Journal of Endocrinological Investigation, 2009

To describe serum and urinary hormones, androgens metabolites and testosterone/epitestosterone ra... more To describe serum and urinary hormones, androgens metabolites and testosterone/epitestosterone ratio profiles after testosterone administration in male hypogonadal volunteers, and to evaluate their possible usefulness in detecting doping with testosterone in treated hypogonadal athletes. Controlled open label design vs placebo; pharmacokinetic study. Ten male volunteers affected by severe hypogonadism (serum testosterone &amp;amp;amp;amp;amp;amp;amp;amp;lt;2.31 ng/ml). Serum and urinary parameters were evaluated, by radioimmunoassay and gas chromatography-mass spectrometry, before and at different time points for 7/3 weeks after a single administration of testosterone enanthate (250 mg) or placebo, respectively. As partially known, testosterone administration increased, with great individual variability, urinary concentrations of glucuronide testosterone, androsterone, etiocholanolone, 5alpha-androstane- 3alpha,17beta-diol, 5beta-androstane-3alpha,17beta-diol, testosterone/ epitestosterone and testosterone/LH ratios; and decreased epitestosterone and 5alpha-androstane-3beta,17beta-diol/5beta-androstane- 3alpha,17beta-diol ratio. Serum testosterone and dihydrotestosterone increased in all volunteers, and concentrations higher than the upper reference limits were observed in many volunteers until 2 weeks after testosterone administration. Whereas the observed prolonged hyperandrogenism partially limited data interpretation, the report ed characteristics of variation of urinary parameters might be used to suspect testosterone misuse in hypogonadal athletes treated with testosterone enanthate. In this sense, while the actual threshold for tes tos terone/epites tos ter one ratio was confirmed to be of reduced usefulness, we suggest a contemporary evaluation of whole urinary androgen metabolites profile and serum androgens, at specific time points after testosterone enanthate administration. Moreover, an adequate tailoring of treatment, to avoid transitory hyperandrogenism, is highly advisable. Further studies on strategies for detecting doping with testosterone in hypogonadal athletes are warranted.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Development and validation of a liquid chromatography–mass spectrometry procedure after solid-phase extraction for detection of 19 doping peptides in human urine

Forensic Toxicology, 2015

A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously ... more A liquid chromatography–tandem mass spectrometry method was developed and used to simultaneously detect 19 small peptide hormones prohibited in sport and their main metabolites in urine after solid-phase extraction. Detection was achieved using a triple-quadrupole mass spectrometric detector coupled with an electrospray ionization interface after chromatographic separation with an octadecyl column based on fused-core particle technology. Sample pretreatment was performed by solid-phase extraction. The extraction procedure was optimized by comparison of different sorbents and washing/elution protocols. The best results were obtained using a mixed-mode weak cation exchange sorbent, two washing steps (ultrapurified water and methanol), and elution using 300 mM ammonium formate in 25 % ammonia/methanol (10/90) or 25 % ammonia/10 % formic acid/methanol (8/12/80). The procedure was validated in terms of sensitivity (lower limits of detection: 0.05–2.0 ng/ml depending on the target analyte), specificity, recovery [>60 %, coefficient of variation (CV) <15 % except for TB500 17–23 fragment, AOD9604, and ARA290 for which recovery was <50 %), ion suppression/enhancement (<35 %), robustness, carryover, stability of the target analytes [stable for at least for 2 days (25 °C, 2 weeks (4 °C), 2 months (−20 °C)], and repeatability of retention times (CV <0.1 %) and relative abundances of the selected ion transitions (CV <15 %). The suitability of the method was confirmed by analyzing spiked and excreted urines, the latter collected after intravenous injection of 0.1 mg of GHRP-2.

Bookmarks Related papers MentionsView impact

Research paper thumbnail of A screening method for the detection of synthetic glucocorticosteroids in human urine by liquid chromatography–mass spectrometry based on class-characteristic fragmentation pathways

Analytical and Bioanalytical Chemistry, Feb 9, 2008

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Detection of synthetic analogues of insulin-like growth factor 1 in different biological fluids by liquid chromatography quadrupole time-of-flight mass spectrometry: comparison of different immunoaffinity protocols

Analytical and Bioanalytical Chemistry

Bookmarks Related papers MentionsView impact

Research paper thumbnail of UHPLC–HRMS Method for the Simultaneous Screening of 235 Drugs in Capillary Blood for Doping Control Purpose: Comparative Evaluation of Volumetric and Non-volumetric Dried Blood Spotting Devices

ACS Omega

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Urinary excretion profile of prednisolone and prednisone after rectal administration: significance in antidoping analysis

Drug Testing and Analysis

Bookmarks Related papers MentionsView impact

Research paper thumbnail of How safe are our internal procedures? Some preliminary experimental evidence on the problem of “lab-oriented” potential masking agents

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Drug-drug interactions and masking effects in sport doping: influence of miconazole administration on the urinary concentrations of endogenous anabolic steroids

Forensic Toxicology, 2016

Bookmarks Related papers MentionsView impact

Research paper thumbnail of Human hepatoma cell lines on gas foaming templated alginate scaffolds for in vitro drug-drug interaction and metabolism studies

Toxicology in vitro : an international journal published in association with BIBRA, Jan 6, 2015

Liver in vitro systems that allow reliable prediction of major human in vivo metabolic pathways h... more Liver in vitro systems that allow reliable prediction of major human in vivo metabolic pathways have a significant impact in drug screening and drug metabolism research. In the present study, a novel porous scaffold composed of alginate was prepared by employing a gas-in-liquid foaming approach. Galactose residues were introduced on scaffold surfaces to promote cell adhesion and to enhance liver specific functions of the entrapped HepG2/C3A cells. Hepatoma cells in the gal-alginate scaffold showed higher levels of liver specific products (albumin and urea) and were more responsive to specific inducers (e.g. dexamethasone) and inhibitors (e.g. ketoconazole) of the CYP3A4 system than in conventional monolayer culture. HepG2/C3A cells were also more efficient in terms of rapid elimination of testosterone, used as a model substance, at rates comparable to those of in vivo excretion. In addition, an improvement in metabolism of testosterone, in terms of phase II metabolite formation, was...

Bookmarks Related papers MentionsView impact