Synbiotic intervention reverses alcohol drinking-induced cognitive deficits in adolescent male mice by modulating the microbiota-gut-brain axis.

Adolescence is characterized by an increased vulnerability to substance abuse, including alcohol consumption. We investigated the effects of a synbiotic intervention on disruptions of the microbiota-gut-brain axis induced by a drinking in the dark model of intermittent alcohol exposure in adolescent mice. We found that alcohol drinking induced specific shifts in gut microbiota, namely it increased Erysipelotrichaceae and reduced fecal butyric and isovaleric acids.

Detection of Oral Testosterone Undecanoate Administration in UGT2B17 del/del and del/ins Individuals. Part II: Urinary Endogenous Steroid Sulfate Markers.

The detection of endogenous anabolic androgenic steroids misuse in Asian population using the Steroidal Module of the Athlete Biological Passport (ABP) is a challenge due to the high prevalence of UGT2B17 gene deletion polymorphism with low levels of testosterone (T) glucuronide. In this study, the capabilities of different approaches based on urine analysis for the detection of oral T undecanoate administration were evaluated in 13 Asian volunteers, including 11 subjects with del/del genotype and 2 subjects with del/ins genotype. In the first part of the work, the effect on the urinary steroid profile (SP) and on the isotope ratio mass spectrometry markers was evaluated.

Detection of Oral Testosterone Undecanoate Administration in UGT2B17 del/del and del/ins Individuals. Part I: Urinary Steroid Profile and IRMS Markers.

The detection of endogenous anabolic androgenic steroids misuse in Asian population using the Steroidal Module of the Athlete Biological Passport (ABP) is a challenge due to the high prevalence of UGT2B17 gene deletion polymorphism and low levels of testosterone (T) glucuronide. In this study, the capabilities of different approaches based on urine analysis for detecting oral T undecanoate administration were evaluated in 13 Asian volunteers, including 11 subjects with del/del genotype and 2 subjects with del/ins genotype. In this part of the work, the effect on the urinary steroid profile (SP) and the isotope ratio mass spectrometry (IRMS) markers were studied.

Towards the non-invasive determination of estradiol levels: Development and validation of an LC-MS/MS assay for quantification of salivary estradiol at sub-pg/mL level.

Background: Estradiol (E2) is a female sex hormone involved in several biological processes. Although E2 levels are commonly measured in blood samples, the use of non-invasive techniques (e.g.

Achieving routine application of dried blood spots for erythropoietin receptor agonist analysis in doping control: low-volume single-spot detection at minimum required performance level.

Erythropoietin receptor agonists (ERAs) are substances prohibited in sports and currently monitored in urine and blood. There is a great interest in new matrices like dried blood spots (DBSs). A direct method for the detection of ERAs in DBSs using one single spot of 25 μl has been optimized and validated.

Effects of structural characteristics of (un)conjugated steroid metabolites in their collision cross section value.

In this work, the collision cross section (CCS) value of 103 steroids (including unconjugated metabolites and phase II metabolites conjugated with sulfate and glucuronide groups) was determined by liquid chromatography coupled to traveling wave ion mobility spectrometry (LC-TWIMS). A time of flight (QTOF) mass analyzer was used to perform the analytes determination at high-resolution mass spectrometry. An electrospray ionization source (ESI) was used to generate [M+H], [M + NH] and/or [M - H] ions.

Potential of desorption electrospray ionization and paper spray ionization with high-resolution mass spectrometry for the screening of sports doping agents in urine.

In this work, desorption electrospray ionization and paper spray ionization both with high-resolution mass spectrometry (DESI-HRMS and PSI-HRMS) were explored for the fast and direct analysis of stimulants and diuretics in urine samples. The analysis was performed at a resolution of 70 000 FWHM (/ 200) using a quadrupole-Orbitrap mass spectrometer in full scan acquisition mode, detecting stimulants and diuretics in positive and negative ion modes, respectively. The most critical parameters affecting the desorption and ionization efficiencies of compounds were optimized, paying particular attention to the optimization of the spray solvent for PSI-HRMS analysis and to the selection of the DESI sample substrate.

Elimination profiles of prednisone and prednisolone after different administration routes: Evaluation of the reporting level and washout periods to ensure safe therapeutic administrations.

Prednisolone (PRED) and prednisone (PSONE) are prohibited in sports competitions when administered by systemic routes, and they are allowed by other routes for therapeutic purposes. There is no restriction of use in out-of-competition periods. The present study aimed to evaluate the urinary excretion of PRED, PSONE, and their most important metabolites after systemic and nonsystemic treatments in order to verify the suitability of the current reporting level of 30 ng/ml used to distinguish allowed and prohibited administrations and to establish washout periods for oral treatments performed in out-of-competition periods.

Comparison of magnetic bead surface functionalities for the immunopurification of growth hormone-releasing hormones prior to liquid chromatography-high resolution mass spectrometry.

Growth hormone-releasing hormone and its analogues sermorelin, tesamorelin and CJC-1295 are included in the prohibited list of the World Antidoping Agency. These target peptides are found at very low concentrations in urine (at the pg/mL level). For this reason, hyphenated enrichment and purification steps prior to mass spectrometric detection are required.

Analysis of hydroxylated phenylalkylamine stimulants in urine by GC-APPI-HRMS.

A gas chromatography-atmospheric pressure photoionization-high-resolution mass spectrometry (GC-APPI-HRMS) method was developed for the determination of eight phenylalkylamine stimulants in urine samples. Spiked urine samples were hydrolyzed, processed by solid-phase extraction, and derivatized before analysis. Two derivatization reactions were studied: the formation of trimethylsilyl (TMS) derivatives with N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA) and trimethylsilyl/trifluoroacetyl (TMS/TFA) derivatives with MSTFA and N-methyl-bis (trifluoroacetamide) (MBTFA) as derivatization reagents.

Elimination profiles of betamethasone after different administration routes: Evaluation of the reporting level and washout periods to ensure safe therapeutic administrations.

Betamethasone (BET) is prohibited in sports competitions when administered by systemic routes, and it is allowed by other routes for therapeutic purposes. In out-of-competition periods, there is no restriction of use. The present work aimed to assess the urinary excretion of BET and its metabolites after allowed and prohibited administrations to verify the suitability of the current reporting level of 30 ng/ml used to distinguish allowed and prohibited administrations and to establish washout periods for oral and intramuscular (IM) administrations when out-of-competition treatments are needed.

Additional studies on triamcinolone acetonide use and misuse in sports: Elimination profile after intranasal and high-dose intramuscular administrations.

Triamcinolone acetonide (TA) is a glucocorticoid (GC) widely used in sports medicine. GCs are prohibited in sports competitions by oral, intramuscular (IM), intravenous and rectal administrations, and they are allowed by other routes considered of local action such as intranasal administration (INT). We examined the urinary profiles of TA and its metabolites after INT and high-dose IM administrations.

Evaluation of sulfate metabolites as markers of intramuscular testosterone administration in Caucasian and Asian populations.

The introduction of alternative markers to the steroid profile can be an effective approach to improving the screening capabilities for the detection of testosterone (T) misuse. In this work, endogenous steroid sulfates were evaluated as potential markers to detect intramuscular (IM) T administration. Fourteen sulfate metabolites were quantified using mixed-mode solid-phase extraction and analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Sulfate metabolites improve retrospectivity after oral testosterone administration.

The detection of testosterone (T) misuse is performed using the steroid profile that includes concentrations of T and related metabolites excreted free and glucuronoconjugated, and the ratios between them. In this work, the usefulness of 14 endogenous steroid sulfates to improve the detection capabilities of oral T administration has been evaluated. Quantitation of the sulfate metabolites was performed using solid-phase extraction and analysis by liquid chromatography-tandem mass spectrometry.

Direct quantitation of endogenous steroid sulfates in human urine by liquid chromatography-electrospray tandem mass spectrometry.

A method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the direct quantitation of endogenous steroid sulfates has been developed to be able to evaluate these metabolites as biomarkers to detect the misuse of endogenous androgenic anabolic steroids in sports. For sample preparation, a mixed-mode solid-phase extraction was optimized to eliminate the glucuronide fraction in the washing step thus obtaining only the sulfate fraction. Chromatographic separation was optimized to achieve adequate resolution between isomers.

Ion-molecule adduct formation in tandem mass spectrometry.

Nowadays most LC-MS methods rely on tandem mass spectrometry not only for quantitation and confirmation of compounds by multiple reaction monitoring (MRM), but also for the identification of unknowns from their product ion spectra. However, gas-phase reactions between charged and neutral species inside the mass analyzer can occur, yielding product ions at m/z values higher than that of the precursor ion, or at m/z values difficult to explain by logical losses, which complicate mass spectral interpretation. In this work, the formation of adduct ions in the mass analyzer was studied using several mass spectrometers with different mass analyzers (ion trap, triple quadrupole, and quadrupole-Orbitrap).

Wide-range screening of psychoactive substances by FIA-HRMS: identification strategies.

Recreational drugs (illicit drugs, human and veterinary medicines, legal highs, etc.) often contain lacing agents and adulterants which are not related to the main active ingredient. Serious side effects and even the death of the consumer have been related to the consumption of mixtures of psychoactive substances and/or adulterants, so it is important to know the actual composition of recreational drugs.

Determination of polyphenolic profiles by liquid chromatography-electrospray-tandem mass spectrometry for the authentication of fruit extracts.

Liquid chromatography-electrospray-tandem mass spectrometry (LC-ESI-MS/MS) was applied to the analysis and authentication of fruit-based products and fruit-based pharmaceutical preparations. A Kinetex C18 reversed-phase column under gradient elution with 0.1 % formic acid aqueous solution and methanol mobile phases was used for the simultaneous determination of 26 polyphenols, allowing an acceptable separation in less than 22 min.

Mixed-mode liquid chromatography coupled to tandem mass spectrometry for the analysis of aminoglycosides in meat.

A novel LC-MS/MS method has been developed for the determination of 13 aminoglycoside antibiotics in meat products. Among the chromatographic columns tested, the mixed-mode Obelisc R provided the best performance. Electrospray has been used for the coupling of the LC and the effect of temperature on the ionization has been evaluated.

Survey of the occurrence of pharmaceuticals in Spanish finished drinking waters.

Fifty samples of finished drinking waters (FDWs) from Spain covering 12 million inhabitants were tested for 53 pharmaceuticals pertaining to 12 different Anatomical Therapeutic Chemical (ATC) classification system codes. The studied compounds are a combination of most commonly consumed pharmaceuticals with other barely reported in the literature. Five compounds, azithromycin, clarithromycin, erythromycin, sulfamethoxazole, and ibuprofen were tentatively identified by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in some samples (2 to 15 %), but only ibuprofen and azithromycin could be confirmed when analyzed by liquid chromatography-high-resolution mass spectrometry (LC-HRMS) with a quadrupole-Orbitrap instrument.

Atmospheric pressure ionization-tandem mass spectrometry of the phenicol drug family.

In this work, the mass spectrometry behaviour of the veterinary drug family of phenicols, including chloramphenicol (CAP) and its related compounds thiamphenicol (TAP), florfenicol (FF) and FF amine (FFA), was studied. Several atmospheric pressure ionization sources, electrospray (ESI), atmospheric pressure chemical ionization and atmospheric pressure photoionization were compared. In all atmospheric pressure ionization sources, CAP, TAP and FF were ionized in both positive and negative modes; while for the metabolite FFA, only positive ionization was possible.

Ultra-high performance liquid chromatography-tandem mass spectrometry for the analysis of phenicol drugs and florfenicol-amine in foods.

A method based on ultra-high performance liquid chromatography (UHPLC) is proposed for the determination of chloramphenicol (CAP), its related compounds, thiamphenicol (TAP) and florfenicol (FF), and the polar metabolite florfenicol-amine (FFA), in animal-derived foods (chicken and pork meat, fish, prawns and honey). For the retention of FFA and its simultaneous analysis with the parent compounds a phenyl-hexyl column is proposed. A fast separation is achieved in less than 2 minutes using a methanol : acetic acid-ammonium acetate buffer (5 mM, pH 5) and gradient elution.

Overview of the structure of all-AT oligonucleotides: organization in helices and packing interactions.

We present the crystalline organization of 33 all-AT deoxyoligonucleotide duplexes, studied by x-ray diffraction. Most of them have very similar structures, with Watson-Crick basepairs and a standard average twist close to 36 degrees. The molecules are organized as parallel columns of stacked duplexes in a helical arrangement.