Name: tlc separation by polarity
Uploaded: 2020-03-26T07:22:28.000Z
Duration: 3 min 7 s
Channel: JoVE

A cromatografia em camada fina, ou TLC, é uma técnica usada para separar compostos orgânicos com base nas diferenças em suas polaridades. Existem três componentes de um sistema TLC: a fase estacionária, o soluto e o solvente de revelação.
A fase estacionária é a superfície na qual os compostos serão separados, que geralmente é composta de sílica gel extremamente polar. Os compostos ou misturas que estão sendo analisados são os solutos. Soluções diluídas dos solutos são localizadas em uma borda da placa usando capilares finos.
O fundo da placa é então imerso no solvente de revelação, que geralmente é um solvente orgânico menos polar do que a fase estacionária. O solvente sobe pela placa por ação capilar. À medida que o solvente passa pelos pontos de soluto, ele carrega um pouco de cada soluto com ele. Esta mistura soluto-solvente é a fase móvel.
O topo da fase móvel é chamado de frente de solvente. À medida que a fase móvel sobe pela placa, as moléculas de soluto e solvente aderem temporariamente à fase estacionária com base em sua polaridade.
Mais solutos polares são fortemente atraídos para a fase estacionária, então eles grudam com mais frequência e permanecem presos por um tempo. Menos solutos polares aderem com menos frequência e não ficam presos por muito tempo. Isso significa que menos solutos polares passam mais tempo na fase móvel do que mais solutos polares. Em outras palavras, menos solutos polares viajam mais rápido do que mais solutos polares.
Para obter uma boa separação entre os compostos, o solvente deve ser polar o suficiente para mover o soluto mais polar. Aqui estão alguns solventes comuns em ordem crescente de polaridade. Às vezes, dois solventes são combinados para atingir a polaridade desejada.
Como a maioria dos compostos orgânicos são brancos ou incolores, as placas TLC geralmente contêm um composto fluorescente. Sob luz ultravioleta, os compostos orgânicos aparecerão como manchas escuras na placa brilhante.
Uma vez que os compostos são visualizados, você pode medir a distância que eles percorreram e calcular o fator de retardo, ouR f, para cada um. Este valor é a distância percorrida pelo soluto dividida pela distância total percorrida pela fase móvel.
Se o solvente for escolhido corretamente, cada soluto terá um Rf distintamente diferente. Isso nos permite identificar os compostos componentes em misturas, combinando os valores de Rf com os compostos conhecidos.
Neste laboratório, você usará o TLC para determinar o Rf de vários compostos conhecidos e, em seguida, usará esses valores de Rf para determinar a identidade de um composto desconhecido.

cromatografia em camada fina

cromatografia
A cromatografia é uma técnica usada em química orgânica para separar compostos em uma mistura com base em suas diferenças de solubilidade entre duas fases diferentes. O conceito é semelhante à extração líquido-líquido, exceto na cromatografia, as duas fases consistem na fase estacionária e na fase móvel. A fase estacionária é um sólido - normalmente um grânulo de hidrogel em microescala - enquanto a fase móvel é um solvente transportador.
Na cromatografia tradicional, a fase estacionária é compactada em uma coluna vertical e a mistura de solução introduzida no topo da coluna. À medida que a mistura flui pela fase estacionária, os compostos se dividem entre a fase estacionária e a fase móvel com base em suas estruturas e polaridades, formando bandas discretas. Os solutos que interagem fracamente com a fase estacionária movem-se mais rapidamente através da coluna e saem - ou eluem - primeiro. Os solutos que interagem mais fortemente com a fase estacionária movem-se lentamente através da coluna e eluem mais tarde. As bandas podem ser coletadas individualmente para isolar e purificar compostos em uma mistura.
Existem vários tipos de cromatografia, cada uma aproveitando uma propriedade química diferente para obter a separação. Por exemplo, na cromatografia de troca iônica, os grânulos de fase estacionária podem ser carregados positiva ou negativamente, atraindo moléculas apenas com carga oposta. Na cromatografia de exclusão de tamanho, a fase estacionária porosa é composta de grânulos como polímeros de agarose ou dextrano. Moléculas menores podem entrar nos poros mais facilmente, enquanto as moléculas maiores fluem pelos poros e eluem mais rapidamente.
<h4>Cromatografia em camada fina</h4>
A cromatografia em camada fina (TLC) é um tipo de técnica de cromatografia que separa compostos com base em sua polaridade. Como a cromatografia tradicional, existem três componentes de um sistema TLC: a fase estacionária, a fase móvel e o soluto. No entanto, ao contrário da cromatografia tradicional, a fase estacionária é disposta em uma camada fina em uma placa, em vez de compactada em uma coluna. A TLC geralmente usa gel de sílica polar, uma forma de dióxido de silício, como fase estacionária. A fase estacionária forma ligações de hidrogênio devido aos grupos OH em sua superfície.
Primeiro, uma linha de partida é desenhada na parte inferior da placa TLC usando um lápis. Os compostos ou misturas que estão sendo analisados são vistos na linha de partida usando um capilar fino. Em seguida, o fundo da placa é imerso na fase móvel, que geralmente é um solvente orgânico menos polar que a fase estacionária. O solvente sobe pela placa por ação capilar, passando pelos pontos de soluto e carregando parte de cada componente com ele.
À medida que o solvente sobe pela placa, os componentes se dividem na fase móvel ou na fase estacionária. Se o componente for polar, ele interage mais com a fase estacionária polar. Ele viaja lentamente e se move apenas uma curta distância na placa TLC. Se o componente da amostra for menos polar - e mais solúvel na fase móvel do que na fase estacionária - ele interage mais com a fase móvel e viaja mais longe na placa TLC. A extensão da polaridade do componente e da fase móvel são essenciais para entender e prever a separação.
As placas TLC normalmente contêm um corante fluorescente reativo a UV que brilha sob uma fonte UV de 254 nanômetros. Portanto, as placas de TLC podem ser analisadas observando-as sob luz UV. Os compostos dentro da placa TLC, como os solutos de interesse, aparecerão como manchas escuras em comparação com um fundo verde. Ao circular as manchas com um lápis de grafite, a distância percorrida pelos compostos em relação à frente do solvente pode ser medida. A mancha do composto orgânico, se não for fluorescente, mascara a fluorescência da placa e aparece como uma mancha escura. Alguns compostos orgânicos são ativos para UV e emitem luz quando expostos à luz UV. Estes são tipicamente compostos conjugados, ou seja, aqueles com ligações duplas e simples alternadas, e podem ser identificados pelo comprimento de onda emitido.
<h4>Fator de retardo</h4>
Ao analisar o fator de retardo (Rf) de um componente com um solvente específico, um soluto desconhecido pode ser determinado usando TLC. O fator de retardo é a razão entre a distância percorrida por um componente e a distância percorrida pela fase móvel.
<img src="/CDNSource/lm/labs/62/62_Concepts_1.jpg">
A distância percorrida pelo soluto é medida da linha de partida até o ponto central do ponto, e a distância percorrida pela fase móvel é medida da mesma linha de partida até a frente do solvente. O fator de retardo de um composto depende da fase móvel usada. O fator de retardo é grande para compostos que são altamente apolares com uma fase móvel apolar. Baixos valores de fator de retardo são observados para componentes polares com uma fase móvel não polar.
No caso de uma fase móvel altamente apolar, alguns componentes polares podem não se mover. Isso resulta em um fator de retardo extremamente baixo e uma separação insuficiente. Uma fase móvel altamente polar faz com que o composto se mova com o solvente e produz um fator de retardo extremamente alto. Isso resulta em muito pouca separação entre os componentes.
Para que a separação seja eficaz, os fatores de retardo dos componentes devem estar separados por cerca de 0,3 a 0,7. Para encontrar a fase móvel eficiente, tentativa e erro são empregados. Muitas vezes, uma mistura de dois solventes se mostra mais eficaz.
<h4>Referências</h4>
<ol>
	<li style="font-size:17px">Harris, D.C. (2015). Análise Química Quantitativa. Nova York, NY: W.H. Freeman and Company.</li>
</ol>

Cromatografia em camada fina

Cromatografia
A cromatografia é uma técnica usada em química orgânica para separar compostos em uma mistura com base em suas diferenças de solubilidade entre ...

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Thin-Layer Chromatography

tlc separation by polarity

Thin-layer chromatography (TLC) separates compounds in a mixture by polarity. It uses the same basic parts as other chromatography methods: a stationary phase, a mobile phase, and the solute. In TLC, the stationary phase is a thin coating on a plate instead of material packed into a column.

The stationary phase in most TLC systems is polar silica gel, a form of silicon dioxide. Its surface has OH groups that can form hydrogen bonds. The sample is first spotted on a pencil start line with a thin capillary. Then the bottom of the plate is placed in the mobile phase, which is usually an organic solvent that is less polar than the stationary phase.

As the solvent moves up the plate by capillary action, it carries the sample components with it. Each compound then partitions between the mobile phase and the stationary phase. A polar component interacts more with the polar stationary phase, so it moves more slowly and stays near the start. A less polar component is more soluble in the mobile phase, so it travels farther up the plate.

TLC plates often include a UV-reactive fluorescent dye that glows under 254 nm UV light. The compounds appear as dark spots against the green background, and the spots can be circled with a graphite pencil so their movement can be measured. Some organic compounds are UV-active and emit light when exposed to UV light. These are often conjugated compounds, which have alternating double and single bonds, and they can be identified by the wavelength they emit.

The retardation factor, or Rf, helps identify an unknown solute from TLC results. It is the ratio of the distance traveled by the compound to the distance traveled by the solvent front. The compound distance is measured from the start line to the center of the spot, and the solvent distance is measured from the same start line to the solvent front. Rf depends on the mobile phase used.

Rf values are high for highly non-polar compounds with a non-polar mobile phase, and low for polar compounds with a non-polar mobile phase. If the mobile phase is too non-polar, some polar compounds may not move at all. If it is too polar, the compound may move with the solvent and the separation may be poor. For an effective separation, the Rf values of the components should be about 0.3 to 0.7 apart. Choosing the best mobile phase usually takes trial and error, and a mixture of two solvents often works well.

TLC Separation by Polarity

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TLC Separation by Polarity

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