Publications by authors named "Bruno Spinosa De Martinis"

28 Publications

  • Page 1 of 1

Analysis of urinary VOCs using mass spectrometric methods to diagnose cancer: A review.

Clin Mass Spectrom 2020 Nov 31;18:27-37. Epub 2020 Oct 31.

Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto - Universidade de São Paulo. Av., Bandeirantes, 3900, Ribeirão Preto, SP 14040-900, Brazil.

The development of non-invasive screening techniques for early cancer detection is one of the greatest scientific challenges of the 21st century. One promising emerging method is the analysis of volatile organic compounds (VOCs). VOCs are low molecular weight substances generated as final products of cellular metabolism and emitted through a variety of biological matrices, such as breath, blood, saliva and urine. Urine stands out for its non-invasive nature, availability in large volumes, and the high concentration of VOCs in the kidneys. This review provides an overview of the available data on urinary VOCs that have been investigated in cancer-focused clinical studies using mass spectrometric (MS) techniques. A literature search was conducted in ScienceDirect, Pubmed and Web of Science, using the keywords "Urinary VOCs", "VOCs biomarkers" and "Volatile cancer biomarkers" in combination with the term "Mass spectrometry". Only studies in English published between January 2011 and May 2020 were selected. The three most evaluated types of cancers in the reviewed studies were lung, breast and prostate, and the most frequently identified urinary VOC biomarkers were hexanal, dimethyl disulfide and phenol; with the latter seeming to be closely related to breast cancer. Additionally, the challenges of analyzing urinary VOCs using MS-based techniques and translation to clinical utility are discussed. The outcome of this review may provide valuable information to future studies regarding cancer urinary VOCs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.clinms.2020.10.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8600992PMC
November 2020

Analysis of Stimulants in Sweat and Urine Using Disposable Pipette Extraction (DPX) and Gas Chromatography Coupled to Mass Spectrometry in the Context of Doping Control.

J Anal Toxicol 2021 Nov 18. Epub 2021 Nov 18.

Department of Chemistry, Faculty of Philosophy, Sciences and Letters of Ribeirao Preto, University of São Paulo, Av. Bandeirantes 3900, Ribeirão Preto, SP, 14040-901, Brazil.

Urine is initially collected from athletes to screen for the presence of illicit drugs. Sweat is an alternative sample matrix that provides advantages over urine including reduced opportunity for sample adulteration, longer detection-time window, and non-invasive collection. Sweat is suitable for analysis of the parent drug and metabolites. In this study, a method was developed and validated to determine the presence of 13 amphetamine and cocaine related substances and their metabolites in sweat and urine using disposable pipette tips extraction (DPX) by GC-MS. The DPX extraction was performed using 0.1 M HCl and dichloromethane: isopropanol: ammonium hydroxide (78:20:2, v/v/v) followed by derivatization with N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) at 90˚C for 20 min. DPX extraction efficiencies ranged between 65.0% and 96.0% in urine and 68.0% and 101.0% in sweat. Method accuracy was 90.0 to 104.0% in urine and 89.0 to 105.0% in sweat. Intra-assay precision in urine and in sweat were lower than 15.6% and 17.8%, respectively, and inter-assay precision ranged from 4.70% to 15.3% in urine and from 4.05% to 15.4% in sweat. Calibration curves presented a correlation coefficient greater than 0.99 for all analytes in both matrices. The validated method was applied to urine and sweat samples collected from forty professional athletes who knowingly took one or more of the target illicit drugs. Thirteen of 40 athletes were positive for at least one drug. All the drugs detected in the urine were also detected in sweat samples indicating that sweat is a viable matrix for screening or confirmatory drug testing.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jat/bkab116DOI Listing
November 2021

Quantifying ethanol in ethanol-based hand sanitizers by headspace gas chromatography with flame ionization detector (HS-GC/FID).

J AOAC Int 2021 Sep 21. Epub 2021 Sep 21.

Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto-Universidade de São Paulo, Av. Bandeirantes, 3900, Ribeirão Preto-, SP, 14040-900, Brazil.

Background: The COVID-19 pandemic sharply increased the demand for ethanol-based gel hand sanitizers, leading to a shortage of these products. Consequently, regulatory health agencies worldwide have altered their regulatory guidelines, on ethanol quality, to meet this high demand, raising concern about the products quality.

Objective: The aim of this study was to quantify ethanol content and to qualitatively assess common impurities in ethanol-based gel hand sanitizers by headspace gas chromatography with flame ionization detector (HS-GC/FID).

Methods: To quantify the ethanol content, 0.10 g of the sample was weighed in a 20 mL headspace vial and 5 mL of deionized water was added. Regarding the qualitative approach, 0.25 g of the sample was weighed and 4 mL of deionized water and was added. The samples were incubated, and then 400 µL of the headspace was injected into the GC/FID. Forty-eight products purchased in Brazil were analyzed.

Results: Thirteen products presented at least one nonconformity regarding the ethanol quantity. Two samples presented an average ethanol concentration below the lower limit considered effective. Twelve samples presented acetaldehyde or ethyl acetate.

Conclusion: The huge demand for ethanol-based gel hand sanitizers may have impacted their quality. Because concern with proper hand hygiene tends to remain an issue for a long period, more studies about quality control of hand sanitizers will be needed.

Highlights: A simple and fast HS-GC/FID method to quantify ethanol in ethanol-based gel hand sanitizers was developed, validated and applied to commercial samples in Brazil. The regulatory authorities must be more vigilant to ensure that the commercially available products meet the recommended specifications.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jaoacint/qsab121DOI Listing
September 2021

Socio-demographic and psychological features associated with smoking in pregnancy.

J Bras Pneumol 2021 1;47(5):e20210050. Epub 2021 Sep 1.

Departamento de Clínica Médica, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto (SP) Brasil.

Objective: To investigate how social and psychological characteristics differ between pregnant women who smoke and do not smoke. To explore associations between social and psychological features with changes of smoking habits by the end of pregnancy.

Methods: A case-control study was set up. Smokers cases were never-smokers and ex-smokers controls. Pregnant women (n=328) from public prenatal services were interviewed. Socio-demographic data and psychological variables - personality traits, anxiety, depression, perceived stress, maternal fetal-attachment - were measured. Saliva samples were collected to measure cotinine and to check self-informed smoking status. In addition, 66 smokers were also assessed regarding smoking habits by late pregnancy. Smoking status was defined as a dependent variable. Exposure factors were analyzed through odds ratios. Logistic models and contingency tables were employed according to the nature of variables. "Qualitative change in smoking" was defined as a dependent variable for the last evaluation, and a logistic regression model was built.

Results: Lower schooling, higher age, use of alcohol and drugs, living without a partner, and passive smoking showed associations with smoking. Anxiety, depression and perceived stress also exhibited positive association with smoking. Among personality traits, only Neuroticism was associated with smoking. None of the variables were associated with qualitative change in smoking by the end of pregnancy.

Conclusion: Smoking during pregnancy is associated with more unfavorable social conditions. Pregnant women who smoke exhibit more negative psychological states than nonsmokers, including a profile of accentuated Neuroticism. None of the investigated variables could predict changes in smoking during pregnancy.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.36416/1806-3756/e20210050DOI Listing
October 2021

Induced sputum abnormalities in gas station attendants.

Int Arch Occup Environ Health 2021 Aug 25. Epub 2021 Aug 25.

Internal Medicine Department. Medical School of Ribeirão Preto, University of São Paulo, Avenida Bandeirantes 3900, Ribeirao Preto, SP, 14048-900, Brazil.

Purpose: To investigate inflammatory changes in the induced sputum (IS) of gas station attendants (GSAs) at risk of exposure to fuel vapors through inhalation and susceptible to respiratory complaints and impaired lung function.

Methods: Hypertonic saline-IS was collected from 52 GSAs who had never smoked (42 men, age = 35.9 ± 8.9 years) and had no known comorbidities. A group of 22 non-smokers (11 men, age = 30.5 ± 5.1 years) selected from the University staff served as control. The GSAs answered a questionnaire and underwent spirometry and chest tomography. A total of 15 inflammatory biomarkers associated with inflammation, including cytokines, chemokines, and mediators of immunological response, were also measured.

Results: The most common symptoms of GSAs were coughing (42.3%) and dyspnea (59.6%) based on the New York Heart Association (NYHA; Class II) classification. Significant elevations (p < 0.05) in forced vital capacity and forced expiratory volume within the first second were observed in the GSAs relative to those in the controls (GSA vs. control: 99 ± 12% vs. 90 ± 9% and 94 ± 11% vs. 87 ± 10%, respectively). The GSAs had a lower percentage of IS lymphocytes than that in the control group (4.5 ± 5.7% vs. 7.7 ± 9.8%). The GSAs also had significantly lower concentrations of IL-4, IL-5, IL-10, IL-12, IFN-γ, and MIP-1α, but IL-3 levels were higher. No differences were observed in the airway thickness and the amount of emphysema between the GSAs and the controls.

Conclusion: Despite normal lung function and absence of abnormalities on HRCT, GSAs have a higher frequency of respiratory complaints, with evidence of impairment of lymphocytic activity in the airways.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00420-021-01755-2DOI Listing
August 2021

Pre-workout supplements marketed in Brazil: Caffeine quantification and caffeine daily intake assessment.

Drug Test Anal 2021 Apr 9. Epub 2021 Apr 9.

Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.

The stimulating and performance-enhancing properties of caffeine are often explored in one the most consumed types of supplements: the pre-workout supplements (PWS). However, despite the popularity of PWS, previous studies have reported incompatibilities between what is described in their labels and their actual caffeine content. This study aimed to develop, to optimize, and to validate a gas-chromatography coupled to nitrogen-phosphorus detector (GC-NPD) method to quantify caffeine in PWS and to analyze commercial PWS marketed in Brazil to estimate the caffeine daily intake. For this purpose, three different extraction procedures were evaluated: agitation in vortex, shaker, and sonication. Sonication yielded the best extraction results. Next, the parameters' temperature and time were optimized by using central composite rotatable design (CCRD) and response surface methodology, which revealed the optimal values of 70°C and 10 min. The method was validated and applied to quantify caffeine in 52 PWS. From the 36 PWS labels that specified the caffeine amount, seven (19%) presented more than 120% of the declared quantity, whereas 15 (42%) contained less than 80% of the labeled caffeine. Additionally, six products presented undeclared caffeine. Considering the label stated doses, five supplements exceeded the safe caffeine daily intake (400 mg). On the basis of these findings, supplement quality control remains an issue that deserves more attention from consumers, manufacturers, and regulatory agencies. Finally, we suggest that PWS consumers be careful of the habit of ingesting caffeine through other sources and avoid ingesting two or more different PWS products in the same day.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/dta.3043DOI Listing
April 2021

Evaluation of salivary VOC profile composition directed towards oral cancer and oral lesion assessment.

Clin Oral Investig 2021 Jul 2;25(7):4415-4430. Epub 2021 Jan 2.

Interdisciplinary Centre of Modern Technologies, Nicolaus Copernicus University in Toruń, 4 Wileńska St., 87-100, Toruń, Poland.

Objectives: Endogenous substances have been analyzed in biological samples in order to be related with metabolic dysfunctions and diseases. The study aimed to investigate profiles of volatile organic compounds (VOCs) from fresh and incubated saliva donated by healthy controls, individuals with oral tissue lesions and with oral cancer, in order to assess case-specific biomarkers of oxidative stress.

Materials And Methods: VOCs were pre-concentrated using headspace-solid phase microextraction and analyzed using gas chromatography-mass spectrometry. Then, VOCs positively modulated by incubation process were subtracted, yielding profiles with selected features. Principal component analysis and hierarchical cluster analysis were used to inspect data distribution, while univariate statistics was applied to indicate potential markers of oral cancer. Machine learning algorithm was implemented, aiming multiclass prediction.

Results: The removal of bacterial contribution to VOC profiles allowed the obtaining of more specific case-related patterns. Artificial neural network model included 9 most relevant compounds (1-octen-3-ol, hexanoic acid, E-2-octenal, heptanoic acid, octanoic acid, E-2-nonenal, nonanoic acid, 2,4-decadienal and 9-undecenoic acid). Model performance was assessed using 10-fold cross validation and receiver operating characteristic curves. Obtained overall accuracy was 90%. Oral cancer cases were predicted with 100% of sensitivity and specificity.

Conclusions: The selected VOCs were ascribed to lipid oxidation mechanism and presented potential to differentiate oral cancer from other inflammatory conditions.

Clinical Relevance: These results highlight the importance of interpretation of saliva composition and the clinical value of salivary VOCs. Elucidated metabolic alterations have the potential to aid the early detection of oral cancer and the monitoring of oral lesions.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00784-020-03754-yDOI Listing
July 2021

Application of microextraction techniques in alternative biological matrices with focus on forensic toxicology: a review.

Bioanalysis 2021 Jan 16;13(1):45-64. Epub 2020 Dec 16.

Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto - Universidade de São Paulo. Av. Bandeirantes, 3900, Ribeirão Preto - SP, 14040-900, Brazil.

The interest in alternative biological matrices (e.g., hair and saliva) for forensic toxicology analysis has increased, and recent developments in sample preparation have targeted rapid, cheap, efficient and eco-friendly methods, including microextraction techniques. For this review, we have gathered information about these two hot topics. We discuss the composition, incorporation of analytes and advantages and disadvantages of different biological matrices, and also present the operation principles of the most reported microextraction procedures and their application in forensic toxicology. The outcome of this review may encourage future forensic researches into alternative samples and microextraction techniques.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.4155/bio-2020-0241DOI Listing
January 2021

Psychoactive substances in human breast milk: a review of analytical strategies for their investigation.

Bioanalysis 2020 Sep 7;12(17):1263-1274. Epub 2020 Sep 7.

Departamento de Química da Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, University of São Paulo, Av. Bandeirantes, 3900, Campus da USP, Ribeirão Preto, SP 14040-900, Brazil.

WHO recommends breastfeeding for the first 6 months of newborn's life. Due to its physicochemical properties, breast milk may contain undesirable components originated from mother's feeding, medication and illicit drugs consumption. Some of these substances transferred from bloodstream to milk and delivered to the infant can cause harmful effects. For the last decades, analytical advances enabled the analysis of several substances in milk using different techniques. Thereby, it is possible to evaluate infant's level of exposure to these substances. This review presents the information published in the main scientific dissemination media about psychoactive drugs investigation in human breast milk, involving the sample preparation techniques and chromatographic validated methods developed in the past 10 years.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.4155/bio-2020-0155DOI Listing
September 2020

Investigation of Ayahuasca β-Carboline Alkaloids and Tryptamine in Sweat Samples from Religious Community Participants by GC-MS.

J Anal Toxicol 2020 Jul;44(6):601-609

Department of Chemistry, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, 3900 Av. Bandeirantes, SP 14050-140, Ribeirão Preto, Brazil.

Ayahuasca, a hallucinogenic beverage used in religious rituals in South America, has become a global phenomenon. Its main active components are the β-carbolines alkaloids, harmine (HRM) and harmaline (HRL), as well as the potent hallucinogen N,N-dimethyltryptamine (DMT). Despite its rising consumption, information regarding possible clinical applications and toxicological effects of ayahuasca is still limited. This study presents the first investigation of the use of sweat for the determination of DMT, HRM and HRL in ayahuasca users during a religious ritual. Sweat is an alternative matrix with advantages over many conventional biological samples, mainly because the collection procedure is non-invasive, easy and simple and samples can be collected without disturbing the religious ritual. In the study, solid-phase extraction was performed under basic conditions. Linearity was observed ranging from 20 to 1500 ng/patch with coefficients of determination (R2) higher than 0.99 for all analytes. The results indicated high selectivity for all investigated analytes, with extraction efficiency exceeding 70%, accuracy ranging from 87.5 to 102.4%, intra-assay precision of 1.85-9.44% and inter-assay precision between 3.34 and 9.85%. The limits of detection were 15 ng/patch for HRM and HRL and 10 ng/patch for DMT. The sweat proved to be a viable option to monitor ayahuasca use.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jat/bkz116DOI Listing
July 2020

Investigation of sweat VOC profiles in assessment of cancer biomarkers using HS-GC-MS.

J Breath Res 2020 02 25;14(2):026009. Epub 2020 Feb 25.

Department of Chemistry, Faculty of Philosophy, Sciences and Letters of Ribeirão Preto, University of São Paulo, Ribeirão Preto, CEP 14040-901, Brazil.

Volatile organic compounds (VOCs) have been studied in biological samples in order to be related to the presence of diseases. Sweat can represent substances existing in blood, has less complex composition (compared with other biological matrices) and can be obtained in a non-invasive way. In this work, sweat patches were collected from healthy controls and volunteers with cancer. Static Headspace was used for VOCs extraction, analysis was performed by gas chromatography coupled with mass spectrometry. Principal Components Analysis was used to investigate data distribution. Random Forest was employed to develop classificatory models. Controls and positive cases could be distinguished with maximum sensitivity and specificity (100% of accuracy) in a model based on the incidence of 2-ethyl-1-hexanol, hexanal and octanal. Discrimination between controls, primary tumors and metastasis was achieved using a panel with 11 VOCs. Balanced accuracy of more than 70% was obtained for the classification of a neoplasm site. Total n-aldehydes presented to be strongly correlated with staging of adenocarcinomas, while phenol and 2,6-dimethyl-7-octen-2-ol were correlated with Gleason score. These findings corroborate with the development of accessible screening tools based on VOC analysis and highlight sweat as a promising matrix to be studied in a clinical context for cancer diagnosis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1088/1752-7163/ab5b3cDOI Listing
February 2020

Analysis of 2,4-Dinitrophenol in Postmortem Blood and Urine by Gas Chromatography-Mass Spectrometry: Method Development and Validation and Report of Three Fatalities in the United States.

J Forensic Sci 2020 Jan 20;65(1):183-188. Epub 2019 Aug 20.

Center for Forensic Science Research and Education, 2300 Stratford Avenue, Willow Grove, PA.

2,4-dinitrophenol (2,4-DNP) is a compound used in the early 1900s as a weight-loss drug but later prohibited due to its severe adverse effects, including death. It has however been attracting interest, due to its weight-loss properties, and appears to be re-emerging in forensic casework. As 2,4-DNP is available for use in industry and as a pesticide and easily accessible online, the dissemination of this drug can be fast. The compound exerts its effects through inhibition of ATP synthesis, and corresponding thermogenic energy loss which can be fatal. A method for qualitative and quantitative analysis of 2,4-DNP in blood and urine specimens using GC-MS with hydrogen as carrier gas is described. The method was validated and displayed acceptable performance parameters with linearity (R higher than 0.998), inter-assay imprecision (lower than 10.6%), intra-assay imprecision (lower than 10.7%), and extraction efficiency (92.1%). Stability of 2,4-DNP in blood and urine was studied, and the drug was stable up to 30 days refrigeration or frozen. Six cases in United States suspected to be related to 2,4-DNP were analyzed. Three cases were found to be positive for 2,4-DNP. Concentrations of 2,4-DNP were in the range of 61.6-220 mg/L in urine and <3-114 mg/L in blood. Based on our findings, we suggest that medical examiners and forensic toxicologists be aware of the reappearance of 2,4-DNP, including this compound as a target in death investigations related to weight-loss drugs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/1556-4029.14154DOI Listing
January 2020

A Gas Chromatography-Mass Spectrometry Method for Toxicological Analysis of MDA, MDEA and MDMA in Vitreous Humor Samples from Victims of Car Accidents.

J Anal Toxicol 2018 Nov;42(9):661-666

Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto Universidade de São Paulo, 3900 Bandeirantes Avenue, Ribeirão Preto, SP, Brazil.

Vitreous humor (VH) shows excellent potential as a matrix of choice for postmortem analytical toxicology due to the ease of sampling and low metabolic activity. This study demonstrates a simple and rapid analytical method to identify and quantify 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine and 3,4-methylenedioxy-ethylamphetamine in VH. Samples were collected with a simple eye puncture procedure, followed by liquid-liquid extraction and derivatization with heptafluorobutyric anhydride and analysis via gas chromatography-mass spectrometry. The accuracy of the method ranged 97-103%, intra-assay precision was between 4.54 and 9.14% relative standard deviation (RSD) and interassay precision ranged from 6.92 to 10.59% RSD. Limits of detection and quantification ranged from 1.0 to 2.5 ng/mL and 10 ng/mL, respectively. The validated method was successfully applied to detect methylenedioxyamphetamine derivatives in VH samples collected from victims of fatal car crashes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jat/bky044DOI Listing
November 2018

Understanding alterations on blood and biochemical parameters in athletes that use dietary supplements, steroids and illicit drugs.

Toxicology 2017 Feb 24;376:75-82. Epub 2016 May 24.

Department of Chemistry, Faculty of Philosophy, Sciences and Letters of Ribeirao Preto, University of São Paulo, Brazil.

In recent years it was verified there are an alarming growing number of teenagers and young adults using a combination of dietary supplements (DS) anabolic androgenic steroids (AAS) and drugs of abuse. This practice is used to improve physical fitness and appearance, may cause serious side effects. This article shows the alterations in the hematological and renal function parameters associate with these substances in 40 athletes. This research involved three steps: 1-the administration of a self-completion questionnaire ; 2-the assessment of hematological and biochemical parameters of renal function and; 3-toxicological urinalysis. Hematological and biochemical tests were conducted in an accredited laboratory and the toxicological urinalysis was validated in our laboratory using liquid-liquid extraction (LLE) and gas chromatography-mass spectrometry (GC-MS). The testosterone levels in the participants who consumed steroids increased 20-60% and alterations in serum creatinine, urea and uric reached values of up to 1.9; 60.6 and 7.5mg/dL, respectively. The toxicological urinalysis supports self-reports confirming the use of AAS and recreational drugs, putting at risk the health of those athletes increasing the chances of kidney diseases.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.tox.2016.05.019DOI Listing
February 2017

Ethanol withdrawal induces anxiety-like effects: Role of nitric oxide synthase in the dorsal raphe nucleus of rats.

Alcohol 2016 05 26;52:1-8. Epub 2016 Feb 26.

Laboratório de Neurobiologia do Estresse e da Depressão, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Avenida Bandeirantes, 3900, Cidade Universitária, 14040-901 Ribeirão Preto, SP, Brazil; Instituto de Neurociências e Comportamento (INeC), Avenida Bandeirantes, 3900, 14049-901 Ribeirão Preto, SP, Brazil; Núcleo de Pesquisa em Neurobiologia das Emoções (NUPNE), Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Avenida Bandeirantes, 3900, Cidade Universitária, 14040-900 Ribeirão Preto, SP, Brazil. Electronic address:

Nitric oxide (NO) mediated transmission in the dorsal raphe nucleus (DRN) has been shown to be involved in the modulation of anxiety-like behaviors. We investigated whether inhibition of nitric oxide synthase (NOS) in the DRN would prevent anxiety-like behavior induced by ethanol withdrawal. Male Wistar rats were treated with ethanol 2-6% (v/v) for a period of 21 days. Ethanol withdrawal was induced by abrupt discontinuation of the treatment. Experiments were performed 48 h after ethanol discontinuation. Rats with a guide cannula aimed at the DRN received intra-DRN injections of the non-selective NOS inhibitor NG-nitro-l-arginine methyl ester (l-NAME), selective neuronal NOS (nNOS) inhibitor N(ω)-propyl-l-arginine (NPLA), or selective inhibitor of inducible NOS (iNOS) N-([3-(aminomethyl)phenyl] methyl) ethanimidamidedihydrochloride (1400W). Five minutes later, the animals were tested in the elevated plus maze (EPM). Plasma ethanol levels were determined by gas chromatography. There was a reduction in plasma ethanol levels 48 h after ethanol withdrawal. Rats from the ethanol withdrawal group showed decreased exploration of the open arms of the EPM with no change in the exploration of enclosed arms. Intra-DRN treatment with l-NAME (100 nmoles/0.2 μL) and 1400W (1 nmol/0.2 μL), but not NPLA (10 nmoles/0.2 μL) in the DRN attenuated the decrease in the exploration of the open arms of the EPM induced by ethanol withdrawal. The major new finding of the present study is that iNOS in the DRN plays a role in the anxiety-like behavior induced by ethanol withdrawal.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.alcohol.2016.02.001DOI Listing
May 2016

Disposable pipette tips extraction: Fundamentals, applications and state of the art.

J Sep Sci 2016 Mar 25;39(6):1168-72. Epub 2016 Feb 25.

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brasil.

Disposable pipette tips extraction consists of a solid-phase extraction in which the sorbent is poorly dispersed in a pipette tip, which allows a quick and dynamic contact between the aspirated analyte from the sample and the solid phase. It is a technique used particularly in food and forensic analysis, since it requires a small amount of sample and solvent. This article highlights the principles, advantages and disadvantages of the disposable pipette tips extraction method and reviews recent applications.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/jssc.201500932DOI Listing
March 2016

Predominance of alcohol and illicit drugs among traffic accidents fatalities in an urban area of Brazil.

Traffic Inj Prev 2016 10 18;17(7):663-7. Epub 2016 Feb 18.

d Departamento de Química , Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo , Ribeirão Preto , Brazil.

Objective: The objective of this study was to determine the prevalence of alcohol and illicit drug use among victims of fatal traffic accidents in the Metropolitan Region of Vitória, Brazil, during the period 2011-2012.

Methods: Blood samples were collected and analyzed for the presence of drugs from 391 deceased victims of traffic crashes that occurred in the Metropolitan Region of Vitória, Brazil. The victims included drivers, passengers, and pedestrians. Sociodemographic variables such as age, gender, day of the week, and period of the year in which the accidents occurred were recorded. The analyses were performed by a gas chromatography-flame ionization method for alcohol and by a gas chromatography-mass spectrometry for amphetamines, cocaine, and cannabis.

Results: The results showed that 44.8% (n = 175) of all cases were positive for alcohol and/or illicit drugs. The detection of alcohol and/or drugs was more frequent in young males, aged 17 to 34, whose samples were positive in 46.8% of cases. Small differences among drivers, passengers, and pedestrians were observed (drivers = 45.9%, passengers = 46.4%, and pedestrians = 45.6%). In general, the most prevalent drug was alcohol, with 141 positive cases (36.1%), followed by cocaine, with 47 positive cases (12%). Amphetamines and cannabis had positivity rates of 4.1 and 4.3%, with 16 and 17 positive cases, respectively. The combined use of alcohol and other drugs was found in 36 cases (9.2%). Crack cocaine use was observed in 27.7% of the positive cases for cocaine.

Conclusions: For the effective reduction of traffic accidents related to driving under influence of drugs (DUID), we suggest the intensification of enforcement actions against the use of alcohol by drivers, the definition of which illicit drugs should be surveyed, as well the cutoff values, the promotion of changing legislation to oblige drivers to provide samples for toxicological testing, and the establishment of public information programs and specific actions aimed at young drivers to promote behavioral changes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/15389588.2016.1146824DOI Listing
October 2016

Ketamine induces anxiolytic effects in adult zebrafish: A multivariate statistics approach.

Behav Brain Res 2015 Oct 15;292:537-46. Epub 2015 Jul 15.

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.

Ketamine inappropriate use has been associated with serious consequences for human health. Anesthetic properties of ketamine are well-known, but its side effects are poorly described, including the effects on anxiety. In this context, animal models are a safe way to conduct this neurobehavioral research and zebrafish (Danio rerio) is an interesting model which has several advantages. The validation and interpretation of results of behavioral assays requires a suitable statistical approach, and the use of multivariate statistical methods has been little explored, especially in zebrafish behavioral models. Here, we investigated the anxiolytic-induced effects of ketamine in adult zebrafish, using Light-Dark Test and proposing the Multivariate Statistics methods (PCA, HCA and SIMCA) to analyze the results. In addition, we compared the processing of data to the one carried out by analysis of variance (ANOVA) ketamine produced significant concentration of exposure-dependent anxiolytic effects, increasing time in white area and number of crossings and decreasing latency to first access to white area. Average entry duration behavior resulted in a slight decrease from control to treatment groups, with an observed concentration-dependent increase among the exposed groups. PCA results indicated that two principal components represent 88.74% of all the system information. HCA and PCA results showed a higher similarity among control and treatment groups exposed to lower concentrations of ketamine and among treatment groups exposed to concentrations of 40 and 60 mg L(-1). In SIMCA results, interclasses distances were concentration of exposure-dependent increased and misclassifications and interclasses residues results also support these findings. These findings confirm the anxiolytic potential of ketamine and zebrafish sensibility to this drug. In summary, our study confirms that zebrafish and multivariate statistics data validation are an appropriate and viable behavioral model for the study of psychoactive substances, providing a detailed and reliable analysis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.bbr.2015.07.017DOI Listing
October 2015

Oral fluid as an alternative matrix to determine ethanol for forensic purposes.

Forensic Sci Int 2014 Sep 30;242:117-122. Epub 2014 Jun 30.

Laboratório de Análises Toxicológicas Forenses, Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Brazil.

The present work aimed to evaluate whether it is possible to use oral fluid to monitor alcohol in drivers. In a control experiment the subjects ingested beer with an alcoholic percentage of 4.7%, in an amount that furnished 0.5 g ethanol per kg of body weight. Volunteer's urine, oral fluid, and breath were collected at 10, 30, 60, and 90 min after alcohol intake. Urine and oral fluid were analyzed by gas chromatography with FID (Flame Ionization Detector); breath was analyzed by Alcotest 7410 (Dräger). The absorption profiles correlated well. The Pearson correlation value between samples of oral fluid and urine, and oral fluid and exhaled air, was close to 1, showing that oral fluid is a promising matrix to monitor drivers in traffic or involved in accidents.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.forsciint.2014.06.024DOI Listing
September 2014

A one-step extraction procedure for the screening of cocaine, amphetamines and cannabinoids in postmortem blood samples.

J Anal Toxicol 2014 Jul-Aug;38(6):341-8. Epub 2014 Apr 29.

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP 14050-140, Brazil.

A gas chromatography-mass spectrometric (GC-MS) method was developed and validated for the simultaneous detection and quantification in postmortem whole blood samples of cocaine (COC), amphetamines (AMPs) and cannabis; the main drugs involved in cases of impaired driving in Brazil. The analytes were extracted by solid-phase extraction by means of Bond-Elute Certify cartridges, derivatized with N-methyl-N-(trimethylsilyl)trifluoroacetamide at 80°C for 30 min and analyzed by GC-MS. Linearity ranged from 10 to 500 ng/mL, except for ecgonine methyl ester, for which linearity ranged from 10 to 100 ng/mL. Inter- and intra-day imprecision ranged from 2.8 to 18.4% and from 1.5 to 14.9%, respectively. Accuracy values lay between 86.9 and 104.4%. The limit of quantitation for all drugs was 10 ng/mL and recoveries were >74% for all analytes, except for cannabinoids, which showed poor recovery (∼30%). The developed method was applied to real samples collected from deceased victims due to traffic accidents. These samples were selected according to the results obtained in immunoassay screening on collected urine samples. Five samples were positive for the presence of COC and metabolites, four samples were positive for cannabinoids, six samples were positive for AMPs and two samples were drug negative. Some samples were positive for more than one class of drug. Results obtained from whole blood samples showed good agreement with urine screening. The developed method proved capable of quantifying all three classes of drugs of abuse proposed in this study, through a one-step extraction procedure.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jat/bku039DOI Listing
June 2015

A rapid assay for the simultaneous determination of nicotine, cocaine and metabolites in meconium using disposable pipette extraction and gas chromatography-mass spectrometry (GC-MS).

J Anal Toxicol 2014 Jan-Feb;38(1):31-8. Epub 2013 Nov 22.

1Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP 14050-140, Brazil.

Drug abuse by pregnant women is considered a serious public health problem worldwide. Meconium is the first excretion in newborns and has been used as an alternative matrix to evaluate in utero drug exposure. Solid phase extraction (SPE) is widely employed to prepare and clean up samples in the field of forensic analysis. Most SPE products require large volumes of solvent, which culminates in longer sample processing times and increased cost per sample. Disposable pipette extraction (DPX) tips have been used as an alternative to traditional SPE cartridges. They combine efficient and rapid extraction with reduced solvent consumption. The purpose of this study was to develop and validate a method to determine nicotine, cotinine, cocaine, benzoylecgonine, cocaethylene and methyl ester anhydroecgonine in meconium using DPX and gas chromatography-mass spectrometry (GC-MS). Validation results indicated that extraction efficiency ranged 50-98%, accuracy 92-106%, intra-assay precision 4-12% and inter-assay precision 6-12%. Linear calibration curves resulted in R(2) values >0.99, limits of detection ranged from 2.5 to 15 ng/g and the limit of quantitation from 10 to 20 ng/g. The DPX-GC-MS method was shown to selectively analyze trace concentrations of drugs in meconium samples. Finally, the developed and validated method was applied to 50 meconium samples.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jat/bkt092DOI Listing
September 2014

Simultaneous quantification of cocaine, amphetamines, opiates and cannabinoids in vitreous humor.

J Anal Toxicol 2014 Jan-Feb;38(1):39-45. Epub 2013 Nov 13.

1Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP 14050-140, Brazil.

A GC-MS method for simultaneous analysis of cocaine (COC), amphetamines (AMPs), opiates, cannabinoids and their metabolites in vitreous humor (VH) was developed and fully validated. VH samples were extracted using solid phase extraction and injected into the GC-MS, using a selected ion monitoring mode. Linearity ranged from 10 to 1000 ng/mL; the exception was anhydroecgonine methyl ester (AEME), for which linearity ranged from 10 to 750 ng/mL. Inter-assay imprecision lay from 1.2 to 10.0%, intra-assay imprecision was <10.4% for all the analytes and accuracy ranged from 95.6 to 104.0%. An limit of quantitation for all drugs was 10 ng/mL and recoveries ranged from 70.4 to 100.1% for basic and neutral compounds; the acid compounds had poor recovery--<40%. The validated method was applied to 10 VH samples taken from individuals whose blood had screened positive for drugs of abuse. All the individuals screened positive for COC in the blood (seven samples) also had positive results in VH; COC concentration ranged from 30.81 to 283.97 ng/mL (mean 186.98 ng/mL) and benzoylecgonine concentration ranged from 11.47 to 460.98 ng/mL (mean 133.91 ng/mL). It was also noticed that, in five cases, cocaethylene was detected. AEME was also quantified in one case. The use of AMP detected by blood analysis was confirmed in the VH of one individual (24.31 ng/mL). However, samples taken from three individuals whose blood tested positive for carboxy-tetrahydrocannabinol presented negative results. The results demonstrated that VH is a suitable alternative biological sample to determine COC, AMPs, opiates and their metabolites.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/jat/bkt093DOI Listing
September 2014

Time-course of neuroendocrine changes and its correlation with hypertension induced by ethanol consumption.

Alcohol Alcohol 2013 Jul-Aug;48(4):495-504. Epub 2013 Jun 2.

Department of Physiology, School of Medicine of Ribeirao Preto, University of Sao Paulo, Sao Paulo, Brazil.

Unlabelled: Ethanol (ETOH) consumption has been associated with endocrine and autonomic changes, including the development of hypertension. However, the sequence of pathophysiological events underlying the emergence of this effect is poorly understood.

Aims: This study aimed to establish a time-course correlation between neuroendocrine and cardiovascular changes contributing to the development of hypertension following ETOH consumption.

Methods: Male adult Wistar rats were subjected to the intake of increasing ETOH concentrations in their drinking water (first week: 5%, second week: 10%, third and fourth weeks: 20% v/v).

Results: ETOH consumption decreased plasma and urinary volumes, as well as body weight and fluid intake. Furthermore, plasma osmolality, plasma sodium and urinary osmolality were elevated in the ETOH-treated rats. ETOH intake also induced a progressive increase in the mean arterial pressure (MAP), without affecting heart rate. Initially, this increase in MAP was correlated with increased plasma concentrations of adrenaline and noradrenaline. After the second week of ETOH treatment, plasma catecholamines returned to basal levels, and incremental increases were observed in plasma concentrations of vasopressin (AVP) and angiotensin II (ANG II). Conversely, plasma oxytocin, atrial natriuretic peptide, prolactin and the hypothalamus-pituitary-adrenal axis components were not significantly altered by ETOH.

Conclusions: Taken together, these results suggest that increased sympathetic activity may contribute to the early increase in MAP observed in ETOH-treated rats. However, the maintenance of this effect may be predominantly regulated by the long-term increase in the secretion of other circulating factors, such as AVP and ANG II, the secretion of both hormones being stimulated by the ETOH-induced dehydration.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/alcalc/agt040DOI Listing
January 2014

Determination of cocaine and metabolites in hair by column-switching LC-MS-MS analysis.

Anal Bioanal Chem 2013 Jul 24;405(19):6299-306. Epub 2013 May 24.

Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, São Paulo, Brazil.

A method for rapid, selective, and robust determination of cocaine (CO) and metabolites in 5-mg hair samples was developed and fully validated using a column-switching liquid chromatography-tandem mass spectrometry system (LC-MS-MS). Hair samples were decontaminated, segmented, incubated overnight in diluted HCl, and centrifuged, and the diluted (1:10 with distilled water) extracts were analyzed in positive ionization mode monitoring two reactions per analyte. Quantifier transitions were: m/z 304.2→182.2 for CO, m/z 290.1→168.1 for benzoylecgonine (BE), and m/z 318.2→196.2 for cocaethylene (CE). The lower limit of quantification (LLOQ) was set at 0.05 ng/mg for CO and CE, and 0.012 ng/mg for BE. Imprecision and inaccuracy at LLOQ were lower than 20 % for all analytes. Linearity ranged between 0.05 and 50.0 ng/mg for CO and CE and 0.012 and 12.50 ng/mg for BE. Selectivity, matrix effect, process efficiency, recovery, carryover, cross talk, and autosampler stability were also evaluated during validation. Eighteen real hair samples and five samples from a commercial proficiency testing program were comparatively examined with the proposed multidimensional chromatography coupled with tandem mass spectrometry procedure and our reference gas chromatography coupled to mass spectrometry (GC-MS) method. Compared with our reference GC-MS method, column-switching technique and the high sensitivity of the tandem mass spectrometry detection system allowed to significantly reduce sample amount (×10) with increased sensitivity (×2) and sample throughput (×4), to simplify sample preparation, and to avoid that interfering compounds and ions impaired the ionization and detection of the analytes and deteriorate the performance of the ion source.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00216-013-7046-3DOI Listing
July 2013

Cannabidiol reduces the anxiety induced by simulated public speaking in treatment-naïve social phobia patients.

Neuropsychopharmacology 2011 May 9;36(6):1219-26. Epub 2011 Feb 9.

Department of Neuroscience and Behavior, School of Medicine of Ribeirão Preto, University of São Paulo, SP, Brazil.

Generalized Social Anxiety Disorder (SAD) is one of the most common anxiety conditions with impairment in social life. Cannabidiol (CBD), one major non-psychotomimetic compound of the cannabis sativa plant, has shown anxiolytic effects both in humans and in animals. This preliminary study aimed to compare the effects of a simulation public speaking test (SPST) on healthy control (HC) patients and treatment-naïve SAD patients who received a single dose of CBD or placebo. A total of 24 never-treated patients with SAD were allocated to receive either CBD (600 mg; n=12) or placebo (placebo; n=12) in a double-blind randomized design 1 h and a half before the test. The same number of HC (n=12) performed the SPST without receiving any medication. Each volunteer participated in only one experimental session in a double-blind procedure. Subjective ratings on the Visual Analogue Mood Scale (VAMS) and Negative Self-Statement scale (SSPS-N) and physiological measures (blood pressure, heart rate, and skin conductance) were measured at six different time points during the SPST. The results were submitted to a repeated-measures analysis of variance. Pretreatment with CBD significantly reduced anxiety, cognitive impairment and discomfort in their speech performance, and significantly decreased alert in their anticipatory speech. The placebo group presented higher anxiety, cognitive impairment, discomfort, and alert levels when compared with the control group as assessed with the VAMS. The SSPS-N scores evidenced significant increases during the testing of placebo group that was almost abolished in the CBD group. No significant differences were observed between CBD and HC in SSPS-N scores or in the cognitive impairment, discomfort, and alert factors of VAMS. The increase in anxiety induced by the SPST on subjects with SAD was reduced with the use of CBD, resulting in a similar response as the HC.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/npp.2011.6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3079847PMC
May 2011

Relation between alcohol consumption and traffic violations and accidents in the region of Ribeirão Preto, São Paulo State.

Forensic Sci Int 2011 Apr 20;207(1-3):164-9. Epub 2010 Oct 20.

Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Brazil.

In recent years, alcohol consumption has been considered an important public health problem. Ethanol, the alcohol used in beverages, is a drug that affects the central nervous system (CNS) and impairs driving skills and co-ordination, increasing risk of deaths and injuries derived from crashes and road accidents. Consumption of alcoholic beverages is implicated with premature deaths, injuries and damages caused by motor vehicle crashes, which result in high costs to government and society. Considering that alcohol consumption is the main responsible factor for deaths and disabilities in young people, the aim of this work was to evaluate the prevalence of blood alcohol in offenders and/or fatal and non-fatal victims of traffic occurrences in the region of Ribeirão Preto, São Paulo State, from 2005 to 2007. The results revealed that in 2134 cases investigated, blood alcohol positivity was generally found in young adults, 25-45 years old and male. The study showed the high risk of drinking and driving and the importance in establishing actions of prevention and intervention to promote the reduction in the number of traffic occurrences related to consumption of alcoholic beverages.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.forsciint.2010.09.021DOI Listing
April 2011

Methodology for urinary 8-hydroxy-2'-deoxyguanosine analysis by HPLC with electrochemical detection.

Pharmacol Res 2002 Aug;46(2):129-31

Department of Pathology, Center of Legal Medicine, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Rua Tenente Catão Roxo 2418, Ribeirão Preto SP14051-140, Brazil.

The compound 8-hydroxy-2'-deoxyguanosine (8-OHdG), one of the major oxidative DNA base products, is a commonly used biomarker of oxidative DNA damage. The routine measurement of 8-OHdG in biologic samples, and especially in urine, is inherently a challenging analytical problem due to the low levels of the analyte and the complexity of the sample. In this study, a simple and rapid procedure based on one step solid phase extraction (SPE) followed by high-performance liquid chromatography with electrochemical detection (HPLC-ECD) analysis is described. The methodology proposed resulted in a recovery of 74.5+/-12% for urinary 8-OHdG with a detection limit of 5.0 microg l(-1).
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/s1043-6618(02)00080-4DOI Listing
August 2002

Automated headspace solid-phase microextraction and capillary gas chromatography analysis of ethanol in postmortem specimens.

Forensic Sci Int 2002 Aug;128(3):115-9

Department of Pathology, Faculty of Medicine of Ribeirão Preto, Center of Legal Medicine, University of São Paulo, Rua Tenente Catão Roxo 2418, Ribeirão Preto, São Paulo 14051-140, Brazil.

Solid-phase microextraction (SPME) is a relatively new solventless sample preparation technique that allows simultaneous sampling, extraction, pre-concentration, and introduction of analytes from a sample matrix in a single procedure. This methodology has been used for the analysis of several drugs of forensic toxicology interest including volatile compounds. This paper describes a methodology for analysis of ethanol and other volatile compounds using automatic headspace solid-phase microextraction (HS-SPME) and capillary gas chromatography in postmortem specimens. The methodology was initially developed using standard solutions of acetaldehyde, acetone, methanol, and ethanol. Isobutanol was used as internal standard. Postmortem samples of blood, urine, and vitreous humor were obtained during medico-legal autopsies. To date, there are no published paper regarding alcohol analysis in vitreous humor specimens using HS-SPME and limited literature analyzing blood and urine samples. HS-SPME analysis showed that, under optimized conditions, ethanol and isobutanol (internal standard) were well-separated from other volatile compounds such as acetaldehyde, acetone, and methanol considered to be potential interferents in ethanol analysis. The calibration curves for each volatile compound demonstrated good linearity throughout the concentration range from 0.001 to 1.0 g/dl and the detection limit of ethanol in the studied specimens was approximately 0.0001 g/dl.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/s0379-0738(02)00182-2DOI Listing
August 2002
-->