Chromatography 1 of 26 Boardworks Ltd 2016
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1 Chromatography 1 of 26 Boardworks Ltd 2016
2 Chromatography 2 of 26 Boardworks Ltd 2016
3 What is chromatography? 3 of 26 Boardworks Ltd 2016 Different instrumental methods can be used to analyse and identify the different components in a mixture. The sensitivity, accuracy and speed of these methods are important, particularly when the mass or concentration of a substance is very small. Chromatography is one such method, and is used to separate a mixture into its different components. There are different types of chromatography, including paper and thin layer chromatography.
4 How chromatography works 4 of 26 Boardworks Ltd 2016 In all types of chromatography, two phases are present: stationary phase a solid or liquid mobile phase a liquid or a gas that flows over the stationary phase. The mobile phase acts as a solvent. This means that the substance to be tested dissolves in the mobile phase. If a liquid solvent is used, it can be either aqueous (water) or non-aqueous (not water). Examples of non-aqueous solvents include alcohols.
5 Movement between the phases 5 of 26 Boardworks Ltd 2016 The mobile phase contains the dissolved components of a mixture. It flows up the stationary phase, carrying the different components. This allows the two phases to exchange components. Components in the mobile phase can attach to the stationary phase. However, this attachment is only temporary, which means it slows the movement of the components along the stationary phase.
6 Separating components 6 of 26 Boardworks Ltd 2016 The different components of a mixture interact with both phases to a different extent. If a component is more strongly attracted to the stationary phase, it s movement through the stationary phase will be slowed. Components more attracted to the mobile phase than the stationary phase will stay dissolved for longer. This is the basis for separating the components of a mixture. The separation of components is dependent on the distribution of the components between the two phases.
7 Different rates of travel 7 of 26 Boardworks Ltd 2016 The speed at which the different components of a mixture travel up the stationary phase depends on two factors: the solubility of the component in the solvent (mobile phase) how easily it dissolves the attraction of the component for the stationary phase. The weaker the attraction for the stationary phase, the faster the component travels and the further along the stationary phase it is found.
8 Chromatography: recap activity 8 of 26 Boardworks Ltd 2016
9 Chromatography 9 of 26 Boardworks Ltd 2016
10 Paper chromatography 10 of 26 Boardworks Ltd 2016 Paper chromatography is used to separate and identify components of mixtures. Dots of single dyes are placed alongside a dot of the unknown mixture. The solvent is drawn up the paper by capillary action. As the solvent moves up the paper, the pattern of the single dyes can be compared to that of the mixture. Which dyes are present in this mixture?
11 Which ink do you think? 11 of 26 Boardworks Ltd 2016
12 Thin layer chromatography 12 of 26 Boardworks Ltd 2016 Remember, all types of chromatography involve a stationary phase and a mobile phase. In thin layer chromatography (TLC) the stationary phase is a layer of silica gel fixed onto a plate. The plate can be glass, foil or plastic. The mobile phase is a solvent that travels up the plate, carrying the substances. silica gel glass plate
13 How does TLC work? 13 of 26 Boardworks Ltd 2016 TLC uses the same basic principle as paper chromatography: capillary action draws the solvent up the plate. A dry sample of the mixture of interest is placed in the silica gel. As the solvent moves up the gel, it dissolves the sample and carries it up the plate. The distance each component travels depends on its attraction to the gel. This depends on the interaction between the solvent, component and gel.
14 TLC or paper chromatography? 14 of 26 Boardworks Ltd 2016 Thin layer chromatography has a number of advantages over paper chromatography. The glass plate is rigid, not flexible like paper, so it is easy to control. Separation is more efficient due to the smaller particle size of the stationary phase. TLC results are easily reproducible. Following TLC, the substances in the mixture can be recovered and the glass plates used again.
15 UV and locating agents 15 of 26 Boardworks Ltd 2016 Many substances are white or colourless, and so are not visible on a TLC plate. One way of making colourless substances show up is to use UV light. An alternative method is to use a chemical locating agent which binds to the substance in the spot. A second chemical is then added which reacts with the locating agent to form a coloured compound.
16 Chromatography 16 of 26 Boardworks Ltd 2016
17 R f values 17 of 26 Boardworks Ltd 2016
18 Calculating R f values 18 of 26 Boardworks Ltd 2016
19 Reference materials 19 of 26 Boardworks Ltd 2016 Scientists can compare the R f value of an unknown compound to R f values of known compounds of that type. The R f values of compounds such as amino acids are published in data books. The amino acids present in a mixture can be identified by comparing the R f values calculated from a chromatogram to those in the data book. amino acid alanine serine leucine lysine valine phenylalanine glycine cysteine R f value
20 Pure or impure? 20 of 26 Boardworks Ltd 2016 A mixture consists of more than one component. It can be referred to as impure. An example is the air, which is a mixture of different gases. Many consumer products, such as drugs, must not contain impurities. Drugs are often mixtures of multiple pure substances that have been specifically combined. A pure sample consists of only one element or compound. It has no impurities. It is important to be able to test a product for impurities using methods such as chromatography.
21 Testing for purity 21 of 26 Boardworks Ltd 2016 As well as identifying components in a mixture, chromatography can be used to determine if a substance is pure. A pure sample can easily be visualised as a single spot on the chromatogram. In contrast, an impure sample, composed of different substances, will be seen as multiple spots on a chromatogram. In the above diagram, what might the pure samples be used for? mixture pure samples
22 Changing the solvent 22 of 26 Boardworks Ltd 2016 Changing the solvent can be useful for identifying different components in a mixture. Different solvents separate mixtures differently, producing different patterns of spots. This means that different Rf values are obtained in different solvents. This is useful as some components may not be soluble in the same solvent or may have similar Rf values in one solvent. What effect would using different solvents on a pure sample have?
23 Chromatography: review activity 23 of 26 Boardworks Ltd 2016
24 Chromatography 24 of 26 Boardworks Ltd 2016
25 Glossary 25 of 26 Boardworks Ltd 2016
26 Multiple-choice quiz 26 of 26 Boardworks Ltd 2016
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