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<!DOCTYPE html>
<html lang="prt-br"> <!-- português -->
<head>
<meta charset="utf-8"> <!-- caracteres especiais -->
<meta http-equiv="X-UA-Compatible" content="IE=edge,chrome=1"> <!-- Permite usar o plug-in do Google Chrome que tem um mecanismo rápido de JavaScript ao abrir páginas no navegador -->
<title>FluxPRT</title> <!-- nome do guia Interativo FluxPRT -->
<meta name="description" content="">
<meta name="viewport" content="width=device-width, initial-scale=1.0"> <!-- permite controlar a janela de visualização, reduzindo a página web inteira para caber na tela de celulares e ipads-->
<link rel="apple-touch-icon" href="apple-touch-icon.png"> <!-- Adapta os icones para vários dispositivos da apple-->
<!-- arquivos CSS -->
<link rel="stylesheet" href="css/bootstrap.min.css">
<link rel="stylesheet" href="css/bootstrap-theme.min.css">
<link rel="stylesheet" href="css/fontAwesome.css">
<link rel="stylesheet" href="css/light-box.css">
<link rel="stylesheet" href="css/owl-carousel.css">
<link rel="stylesheet" href="css/templatemo-style.css">
<link href="https://fonts.googleapis.com/css?family=Open+Sans:300,400,600,700,800" rel="stylesheet"> <!-- chamar a familia da fonte OpenSans -->
<script src="js/vendor/modernizr-2.8.3-respond-1.4.2.min.js"></script> <!-- biblioteca de js que detecta os recursos disponíveis no navegador e informa quando o navegador não é compatível -->
</head>
<body>
<header class="nav-down responsive-nav hidden-lg hidden-md">
<button type="button" id="nav-toggle" class="navbar-toggle" data-toggle="collapse" data-target="#main-nav">
<span class="sr-only">Toggle navigation</span> <!-- têcnica usada para acessibilidade-->
<span class="icon-bar"></span>
<span class="icon-bar"></span>
<span class="icon-bar"></span>
</button>
<!-- cabecalho da barra de navegação /navbar-header/ -->
<div id="cabecalho da barra de navegação" class="collapse navbar-collapse"><nav><ul class="nav navbar-nav">
<li><a href="#top">Estrutura</a></li>
<li><a href="#video">Cromatografia</a></li>
<li><a href="#blog">Tipos de Cromatografia</a></li>
<li><a href="#projects">Quociente</a></li>
<li><a href="#contact">e-book</a></li>
</ul></nav>
</div>
</header>
<div class="sidebar-navigation hidde-sm hidden-xs">
<div class="logo">
<a href="index.html"><em>Flux</em>PRT</a>
</div>
<nav>
<ul>
<li>
<a href="#video">
<em>Cromatografia</em>
</a>
</li>
<li>
<a href="#video">
<span class="rect"></span>
<span class="circle"></span>
Fluxo de trabalho
</a>
</li>
</li>
<li>
<a href="#blog">
<span class="rect"></span>
<span class="circle"></span>
Tipos de Cromatografia
</a>
</li>
<li>
<a href="#projects">
<span class="rect"></span>
<span class="circle"></span>
Quociente
</a>
</li>
<li>
<a href="#contact">
<span class="rect"></span>
<span class="circle"></span>
e-book
</a>
</li>
</ul>
</nav>
<ul class="social-icons">
<!-- página inicial da página web -->
<li><a href="index.html"><i class="fa fa-home"></i></a></li>
<!-- github do guia FluxPRT -->
<li><a href="http://cabernet.luar.dcc.ufmg.br:7070/groups/luar" target="_blank"><i class="fa fa-github"></i></a></li>
<!-- webpage do LUAR -->
<li><a href="http://luar.dcc.ufmg.br/" target="_blank"><i class="fa fa-share-alt"></i></a></li>
<!-- email da desenvolvedora -->
<li><a href="mailto:elizabeth.espinoza@ufv.br" target="_blank"><i class="fa fa-envelope"></i></a></li>
</ul>
</div>
<div class="page-content">
<section id="video" class="content-section">
<div class="row"><div class="col-md-12"><div class="section-heading">
<h1>Você está na atividade <em>Cromatografia</em> do FluxPRT</h1>
<p>O Guia Interativo do FluxPRT segue o Fluxo de Trabalho em Proteômica.</p>
</div>
<div class="text-content">
<p>O Guia Interativo do FluxPRT segue o Fluxo de Trabalho em Proteômica apresentado a seguir, clique nas atividades enumeradas mais detalhes e dicas. Aproveite instruções e dicas para tornar sua experiência com o FluxPRT ainda mais produtiva, nesta seção sao apresentadas informacões a respeito do Cromatografia.<br><br></p>
</div>
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</section>
<section id="blog" class="content-section">
<div class="section-heading">
<h1><em>Tipos</em> de<br>Cromatografia</h1><p>Permite a separação e identificação de componentes de uma mistura.<br>Tipos de acordo ao mecanismo de separação:</p>
</div>
<p>A cromatografia é um processo que se baseia na migração dos compostos de uma mistura, os quais apresentam diferentes interações através de duas fases, uma fixa em interação com o soluto (fase estacionária) e uma fase móvel (Carta and Jungbauer, 2010).</p>
<p>A técnica de cromatografia permite que os componentes da mistura sejam separados por interação diferenciada com a resina, com base em propriedades moleculares como: massa molecular, carga elétrica, solubilidade e afinidade (Fekete, Guillarme, Sandra, and Sandra, 2016). Pode se classificar de acordo ao mecanismo de separação (Ettre, 1993):</p>
<br><br>
<div class="section-content"><div class="tabs-content"><div class="wrapper">
<ul class="tabs clearfix" data-tabgroup="first-tab-group">
<li><a href="#tab1" class="active">Exclusão</a></li>
<li><a href="#tab2">Afinidade</a></li>
<li><a href="#tab3">Hidrofóbica</a></li>
<li><a href="#tab4">Troca Iônica</a></li>
<li><a href="#tab5">Fase Reversa</a></li>
</ul>
<section id="first-tab-group" class="tabgroup">
<!-- Exclusão -->
<div id="tab1"><ul><li><div class="item">
<div class="text-content">
<h4>Cromatografia de Gel de Filtração ou de Exclusão Molecular</h4>
<span>Pela massa molecular</span>
<p>Separação pela Massa Molecular das proteínas. Obs: Permite avaliar as interações proteína-proteína.</p>
</div></div></li></ul></div>
<!-- //Exclusão -->
<!-- Afinidade -->
<div id="tab2"><ul><li><div class="item">
<div class="text-content">
<h4>Cromatografia de Afinidade Molecular</h4>
<span>Pela afinidade da proteína</span>
<p>Separação pela afinidade da proteína por um ligante específico unido à fase estacionária da coluna.</p>
</div></div></li></ul></div>
<!-- //Afinidade -->
<!-- Hidrofóbica -->
<div id="tab3"><ul><li><div class="item">
<div class="text-content">
<h4>Cromatografia de Interação Hidrofóbica</h4>
<span>Pelas interações apolares</span>
<p>Separação pela interação de resíduos de aminoácidos apolares das proteínas e uma matriz hidrofóbica.</p>
</div></div></li></ul></div>
<!-- //Hidrofóbica -->
<!-- Troca Iônica -->
<div id="tab4"><ul><li><div class="item">
<div class="text-content">
<h4>Cromatografia de Troca Iônica</h4>
<span>Pela carga das proteínas</span>
<p>Separação pela carga das proteínas, esta se ligam reversivelmente a um suporte com carga oposta. Obs: Vai depender do ponto isoelétrico das proteínas de interesse e do pH da solução.</p>
</div></div></li></ul></div>
<!-- //Troca Iônica -->
<!-- Fase Reversa -->
<div id="tab5"><ul><li><div class="item">
<div class="text-content">
<h4>Cromatografia de Fase Reversa</h4>
<span>Pela afinidade do solvente</span>
<p>Separação por afinidade a um solvente orgânico. Obs: Pode desnaturar a proteína e consequentemente comprometer a atividade biológica e estrutura 3D desta.</p>
</div></div></li></ul></div>
<!-- /Fase Reversa -->
</section></div></div>
</div>
</section>
<section id="projects" class="content-section">
<div class="section-heading">
<h1>Quociente ou<br><em>Taxa de vazão</em></h1>
<p>O FluxPRT foi desenvolvido como um fluxo de trabalho adaptável de proteômica,
<br>Clique nas atividades e aprenda mais.</p>
</div>
<p>Este é o volume da fase móvel que passa pela coluna em tempo unitário, a vazão (<em>F</em>, de <em>flow rate</em>) é medida na saída da coluna, a pressão (<em>p<sub>a</sub></em>) e temperatura ambiente (<em>T<sub>a</sub></em>, em Kelvin).</p><br>
<div class="tabgroup">
<h4>Obtenção da vazão da fase móvel</h4>
<p>Para obter a vazão da fase móvel (<em>F<sub>a</sub></em>) à temperatura ambiente a partir de um medidor de vazão que contém água (e.g. o medidor da bolha de sabão), temos que corrigir o <em>F </em>para condições de gás seco a partir de (5.1), onde <em>p<sub>w</sub></em> é a pressão parcial do vapor de água á temperatura ambiente (Ettre, 1993; McNaught, Wilkinson, et al., 1997). A taxa de vazão da fase móvel (Fc) à temperatura da coluna(Tc, em Kelvin) pode ser obtida a partir de (5.2).</p>
<p><em>F<sub>a</sub> =F(1-(P<sub>w</sub>/P<sub>a</sub>)) </em> (5.1)</p>
<p><em>F<sub>c</sub> =F<sub>a</sub>(T<sub>c</sub>/T<sub>a</sub>) </em> (5.2)</p>
<p>O fator que tem maior efeito na Taxa de vazão é o diâmetro da coluna (não o comprimento desta), pois desempenha um grande papel na dispersão e no volume dos picos. A vazão ótima para qualquer coluna pode ser calculada por estimativa (Merck; Phenomenex; Waters; Sciences), mas para encontrar a taxa de vazão aplicável ao método real usado precisa ser determinada pela execução de amostras e a análise da eficiência de separação.</p>
</div>
<br><br><strong>Referências</strong><br><br>
<p>Giorgio Carta and Alois Jungbauer. Protein Chromatography: Process Development and Scale-Up. John Wiley & Sons, 1 edition, 2010. ISBN 978-3-527-31819-3</p>
<p>Szabolcs Fekete, Davy Guillarme, Pat Sandra, and Koen Sandra. <a href="https://doi.org/10.1021/acs.analchem.5b04561" target="_blank">Chromatographic, electrophoretic, and mass spectrometric methods for the analytical characterization of protein biopharmaceuticals</a>. Analytical Chemistry, 88(1):480–507, 2016. ISSN 0003-2700. doi: 10.1021/acs.analchem.5b04561.</p>
<p>L. S Ettre. <a href="http://www.rsync.iupac.org/publications/pac/1993/pdf/6504%C3%970819.pdf" target="_blank">Nomenclature for chromatography</a>. Pure and applied chemistry, 1993. ISSN 0033-4545.</p>
<p>Alan D McNaught, Andrew Wilkinson, et al. <a href="https://goldbook.iupac.org/terms/view/F02439" target="_blank">IUPAC</a>. Compendium of Chemical Terminology, volume 1669. Blackwell Scientific Publications, Oxford, 1997. ISBN 0-9678550-9-8. doi: 10.1351/goldbook.</p>
<p>Merck. <a href="https://www.sigmaaldrich.com/analytical-chromatography/hplc/method-transfer-calculator.html" target="_blank">Supelco HPLC Calculator for Isocratic or Gradient Method Transfer |Sigma-Aldrich</a>.</p>
<p>Phenomenex. <a href="https://www.phenomenex.com/Tools/preplc-scaleup" target="_blank">Preparative Scaling Calculator | Phenomenex UHPLC, HPLC, SPE and GC</a>.</p>
<p>Waters. <a href="http://www.waters.com/waters/promotionDetail.htm?id=134713448&alias=Alias_prepcalculator_CHEMISTRY&locale=de_CH" target="_blank">Bridging the Performance Gap From Analytical to Prep</a>.</p>
<p>Analytical Sciences. <a href="https://epgl.unige.ch/labs/fanal/hplc_calculator:en" target="_blank">HPLC Calculator</a>.</p>
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