The development of modern mass spectrometry techniques for the analysis of beneficial and toxic bioactive compounds in food

Orientador: Rodrigo Ramos CatharinoTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: A necessidade de métodos analíticos mais rápidos e com alta eficiência é o que move a pesquisa aplicada às áreas de ciência e análise de alimentos. A metabolômica é uma plataforma extremamente versátil, que integra dados estatísticos e experimentais na produção de resultados analíticos de alta confiabilidade. Aliado às técnicas modernas de espectrometria de massas, isso torna este segmento extremamente fértil para a realização de melhorias e desenvolvimento. Este projeto teve como escopo a análise direta de amostras de diversos alimentos com características funcionais, como azeites, amendoim e chocolates, com mínimo preparo de amostra, através de espectrometria de massas convencional (MS) e por imagem (MSI). Foram avaliadas características químicas de sua composição em casos de adulterações, degradações ou contaminações. Para tal finalidade, fontes de inonização como a dessorção a laser assistida por matriz (MALDI) e o spray de elétrons (ESI) foram amplamente exploradas. Toda a elucidação estrutural teve o respaldo de reações de fragmentação e análises por espectrometria de massas de alta resolução (ESI-Orbitrap). Os compostos (marcadores) foram tanto previamente definidos (target analysis) quanto identificados pós-análise (metabolic fingerprinting)Abstract: The urge for faster and highly efficient analytical methods is the drive force in food research and analysis. Metabolomics is a versatile platform that integrates both statistical and experimental data, providing highly reliable analytical results. Along with the modern mass spectrometric techniques, there is a very prominent scenario for the development of new and improved approaches. This project aimed at developing direct analysis methods for samples of various functional food products, such as olive oil, peanuts and chocolate through both conventional mass spectrometry (MS) and mass spectrometry imaging (MSI), with little sample preparation. Chemical characterization, degradation and adulteration processes were abundantly monitored within this scope. Ionization sources, such as matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) were largely explored tools in this context. Structural elucidations were performed with the assistance of high-resolution mass spectrometry (ESI-Orbitrap). Chemical markers were either previously elected for a targeted analysis, or identified after the analytical process, as in metabolic fingerprintingDoutoradoFisiopatologia MédicaDoutor em Ciências2014/00084-2FAPES

Novel "omic" strategies applied to pharmaceutical products

Orientador: Rodrigo Ramos CatharinoTexto em português e inglêsDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: Com a larga distribuição e o crescimento de mercado da indústria de cosméticos, esta deve sempre seguir processos de produção e controle de qualidade altamente eficazes. Ensaios analíticos dessa natureza são realizados utilizando-se técnicas de cromatografia, podendo ou não ser acopladas a análises por espectrometria de massas. Entretanto, esses procedimentos requerem fases extensas de preparo de amostra, englobando desde processos de extração até mesmo a reações de derivatização. Num cenário industrial, isso se reflete em grande demanda de tempo e, portanto, custos adicionais. Tendo em vista esse escopo, apresentamos uma nova plataforma "ômica" ¿ a Cosmetômica, que propõe o uso de metodologias simplificadas e de grande eficácia em termos de preparo de amostra e facilidade na aquisição de dados. Amostras de diferentes tipos de cosméticos são analisadas através da técnica de espectrometria de massas por imagem (MALDI-MSI), qualitativa e quantitativamente, através da integração de dados tanto analíticos quanto estatísticos. É possível controle total de toda a cadeia produtiva, desde a matéria-prima até o produto acabado, sendo esse último o objeto de estudo do presente trabalhoAbstract: With the large distribution and the increasingly promising market of cosmetic industry, highly-qualified productive and quality control processes are mandatory for companies to follow. Analytical procedures for QC are commonly carried out using instrumental approaches such as chromatography ¿ coupled or not with mass spectrometry. However, these procedures require sample preparation steps, which may comprise extraction and even sample derivatization. In an industrial scenario it may reflect in great demand of time, with elevated costs as the main consequence. Within this scope, we present a new "omic" platform ¿ Cosmetomics, which proposes the use of simple and effective methods in terms of sample preparation and readiness of data acquisition. Samples from different types of daily use cosmetics are analyzed by mass spectrometry imaging (MALDI-MSI), both qualitative and quantitatively, through the integration of analytical and statistical data. It is possible, therefore, full control of the whole productive chain, from the raw material up to the finished product, which was the elected sample class of our studyMestradoCiencias BiomedicasMestre em Ciências Médica

In Vitro Evaluation Of Sun Protection Factor And Stability Of Commercial Sunscreens Using Mass Spectrometry

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Sunlight exposure causes several types of injury to humans, especially on the skin; among the most common harmful effects due to ultraviolet (UV) exposure are erythema, pigmentation and lesions in DNA, which may lead to cancer. These long-term effects are minimized with the use of sunscreens, a class of cosmetic products that contains UV filters as the main component in the formulation; such molecules can absorb, reflect or diffuse UV rays, and can be used alone or as a combination to broaden the protection on different wavelengths. Currently, worldwide regulatory agencies define which ingredients and what quantities must be used in each country, and enforce companies to conduct tests that confirm the Sun Protection Factor (SPF) and the UVA (Ultraviolet A) factor. Standard SPF determination tests are currently conducted in vivo, using human subjects. In an industrial mindset, apart from economic and ethical reasons, the introduction of an in vitro method emerges as an interesting alternative by reducing risks associated to UV exposure on tests, as well as providing assertive analytical results. The present work aims to describe a novel methodology for SPF determination directly from sunscreen formulations using the previously described cosmetomics platform and mass spectrometry as the analytical methods of choice. (C) 2015 Elsevier B.V. All rights reserved.9881319Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)INCTConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [processes 2014/00084-2, 2011/50400-0, 2013/20066-6

In vitro evaluation of sun protection factor and stability of commercial sunscreens using mass spectrometry

Sunlight exposure causes several types of injury to humans, especially on the skin; among the most common harmful effects due to ultraviolet (UV) exposure are erythema, pigmentation and lesions in DNA, which may lead to cancer. These long-term effects are minimized with the use of sunscreens, a class of cosmetic products that contains UV filters as the main component in the formulation; such molecules can absorb, reflect or diffuse UV rays, and can be used alone or as a combination to broaden the protection on different wavelengths. Currently, worldwide regulatory agencies define which ingredients and what quantities must be used in each country, and enforce companies to conduct tests that confirm the Sun Protection Factor (SPF) and the UVA (Ultraviolet A) factor. Standard SPF determination tests are currently conducted in vivo, using human subjects. In an industrial mindset, apart from economic and ethical reasons, the introduction of an in vitro method emerges as an interesting alternative by reducing risks associated to UV exposure on tests, as well as providing assertive analytical results. The present work aims to describe a novel methodology for SPF determination directly from sunscreen formulations using the previously described cosmetomics platform and mass spectrometry as the analytical methods of choice9881319COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação2014/00084-2; 2011/50400-0; 2013/20066-6INC

In Vitro Evaluation Of Sun Protection Factor And Stability Of Commercial Sunscreens Using Mass Spectrometry.

Sunlight exposure causes several types of injury to humans, especially on the skin; among the most common harmful effects due to ultraviolet (UV) exposure are erythema, pigmentation and lesions in DNA, which may lead to cancer. These long-term effects are minimized with the use of sunscreens, a class of cosmetic products that contains UV filters as the main component in the formulation; such molecules can absorb, reflect or diffuse UV rays, and can be used alone or as a combination to broaden the protection on different wavelengths. Currently, worldwide regulatory agencies define which ingredients and what quantities must be used in each country, and enforce companies to conduct tests that confirm the Sun Protection Factor (SPF) and the UVA (Ultraviolet A) factor. Standard SPF determination tests are currently conducted in vivo, using human subjects. In an industrial mindset, apart from economic and ethical reasons, the introduction of an in vitro method emerges as an interesting alternative by reducing risks associated to UV exposure on tests, as well as providing assertive analytical results. The present work aims to describe a novel methodology for SPF determination directly from sunscreen formulations using the previously described cosmetomics platform and mass spectrometry as the analytical methods of choice.988C13-1

Screening The Life Cycle Of Schistosoma Mansoni Using High-resolution Mass Spectrometry.

Schistosomiasis is a common tropical disease caused by Schistosoma species Schistosomiasis' pathogenesis is known to vary according to the worms' strain. Moreover, high parasitical virulence is directly related to eggs release and granulomatous inflammation in the host's organs. This virulence might be influenced by different classes of molecules, such as lipids. Therefore, better understanding of the metabolic profile of these organisms is necessary, especially for an increased potential of unraveling strain virulence mechanisms and resistance to existing treatments. In this report, direct-infusion electrospray high-resolution mass spectrometry (ESI(+)-HRMS) along with the lipidomic platform were employed to rapidly characterize and differentiate two Brazilian S. mansoni strains (BH and SE) in three stages of their life cycle: eggs, miracidia and cercariae, with samples from experimental animals (Swiss/SPF mice). Furthermore, urine samples of the infected and uninfected mice were analyzed to assess the possibility of direct diagnosis. All samples were differentiated using multivariate data analysis, PCA, which helped electing markers from distinct lipid classes; phospholipids, diacylglycerols and triacylglycerols, for example, clearly presented different intensities in some stages and strains, as well as in urine samples. This indicates that biochemical characterization of S. mansoni may help narrowing-down the investigation of new therapeutic targets according to strain composition and aggressiveness of disease. Interestingly, lipid profile of infected mice urine varies when compared to control samples, indicating that direct diagnosis of schistosomiasis from urine may be feasible.84562-

In Situ Assessment Of Atorvastatin Impurity Using Maldi Mass Spectrometry Imaging (maldi-msi).

The analysis of impurities and degradation products in pharmaceutical preparations are usually performed by chromatographic techniques such as high-performance liquid chromatography (HPLC). This approach demands extensive analysis time, mostly due to extraction and separation phases. These steps must be carried out in samples in order to adapt them to the requirements of the analytical method of choice. In the present contribution, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was employed to quantify an important degradation product in atorvastatin calcium 80 mg tablets: the atorvastatin lactone. Through the standard of the impurity, it was possible to perform quantitative analysis directly on the drug tablet, using a quick and novel approach, suitable for quality control processes in the pharmaceutical industry.81832-

High-throughput Analysis By Sp-ldi-ms For Fast Identification Of Adulterations In Commercial Balsamic Vinegars.

Balsamic vinegar (BV) is a typical and valuable Italian product, worldwide appreciated thanks to its characteristic flavors and potential health benefits. Several studies have been conducted to assess physicochemical and microbial compositions of BV, as well as its beneficial properties. Due to highly-disseminated claims of antioxidant, antihypertensive and antiglycemic properties, BV is a known target for frauds and adulterations. For that matter, product authentication, certifying its origin (region or country) and thus the processing conditions, is becoming a growing concern. Striving for fraud reduction as well as quality and safety assurance, reliable analytical strategies to rapidly evaluate BV quality are very interesting, also from an economical point of view. This work employs silica plate laser desorption/ionization mass spectrometry (SP-LDI-MS) for fast chemical profiling of commercial BV samples with protected geographical indication (PGI) and identification of its adulterated samples with low-priced vinegars, namely apple, alcohol and red/white wines.83886-9

Comparative metabolomic profiling of women undergoing in vitro fertilization procedures reveals potential infertility-related biomarkers in follicular fluid

Abstract Infertility is a worldwide concern, affecting one in six couples throughout their reproductive period. Therefore, enhancing the clinical tools available to identify the causes of infertility may save time, money, and emotional distress for the involved parties. This study aims to annotate potential biomarkers in follicular fluid that are negatively affecting pregnancy outcomes in women suffering infertility-related diseases such as endometriosis, tuboperitoneal factor, uterine factor, and unexplained infertility, using a metabolomics approach through high-resolution mass spectrometry. Follicular fluid samples collected from women who have the abovementioned diseases and managed to become pregnant after in vitro fertilization procedures [control group (CT)] were metabolically compared with those from women who suffer from the same diseases and could not get pregnant after the same treatment [infertile group (IF)]. Mass spectrometry analysis indicated 10 statistically relevant differential metabolites in the IF group, including phosphatidic acids, phosphatidylethanolamines, phosphatidylcholines, phosphatidylinositol, glucosylceramides, and 1-hydroxyvitamin D3 3-d-glucopyranoside. These metabolites are associated with cell signaling, cell proliferation, inflammation, oncogenesis, and apoptosis, and linked to infertility problems. Our results indicate that understanding the IF’s metabolic profile may result in a faster and more assertive female infertility diagnosis, lowering the costs, and increasing the probability of a positive pregnancy outcome

Skin Imprinting In Silica Plates: A Potential Diagnostic Methodology For Leprosy Using High-resolution Mass Spectrometry

Leprosy is a chronic infectious disease caused by Mycobacterium leprae, which primarily infects macrophages and Schwann cells, affecting skin and peripheral nerves. Clinically, the most common form of identification is through the observation of anesthetic lesions on skin; however, up to 30% of infected patients may not present this clinical manifestation. Currently) the gold standard diagnostic test for leprosy is based on skin lesion biopsy, which is invasive and presents low Sensibility for suspect dases. Therefore, the development of a fast, sensible and noninvasive method that identifies infected patient's would be helpful for assertive diagnosis. The aim of this Work was to identify lipid markers in, leprosy patients directly from skin imprints, using a mass spectrometric analytical strategy. For skin imprint samples, a 1 cm(2) silica plate was gently pressed against the skin of patients or healthy volunteers. Imprinted silica lipids were extracted and submitted to direct-infusion electrospray ionization high-resolution mass spectrometry (ESI-HRMS). All samples were differentiated using a lipidomics-based data workup employing multivariate data analysis, which helped electing different lipid markers, for example, mycobacterial mycolic acids, inflammatory and apoptotic molecules were identified as leprosy patients' markers. Otherwise, phospholipids and gangliosides were pointed as healthy volunteers' skin lipid markers, according to normal skin composition. Results indicate that silica plate skin imprinting associated with ESI-HRMS is a promising fast and sensible leprosy diagnostic method. With a prompt leprosy diagnosis, an early and effective treatment could be feasible and thus the chain of leprosy transmission could be abbreviated.87735853597Natural Science and Engineering Research CouncilCanada Foundation for InnovationAdvanced Electrophoresis Solutions Ltd