The document discusses chromatography techniques used to separate and identify components in mixtures. It describes the basic principles of chromatography, which involve a stationary and mobile phase. Paper chromatography and thin layer chromatography (TLC) are explained as two common techniques. Key points covered include how to read a chromatogram, calculate Rf values to identify unknown substances, and factors that affect separation. The document provides instructions for homework on chromatography questions and calculations.
The document discusses chromatography techniques used to separate and identify components in mixtures. It describes the basic principles of chromatography, which involve a stationary and mobile phase. Paper chromatography and thin layer chromatography (TLC) are explained as two common techniques. Key points covered include how to read a chromatogram, calculate Rf values to identify unknown substances, and factors that affect separation. The document provides instructions for homework on chromatography questions and calculations.
The document discusses chromatography techniques used to separate and identify components in mixtures. It describes the basic principles of chromatography, which involve a stationary and mobile phase. Paper chromatography and thin layer chromatography (TLC) are explained as two common techniques. Key points covered include how to read a chromatogram, calculate Rf values to identify unknown substances, and factors that affect separation. The document provides instructions for homework on chromatography questions and calculations.
The document discusses chromatography techniques used to separate and identify components in mixtures. It describes the basic principles of chromatography, which involve a stationary and mobile phase. Paper chromatography and thin layer chromatography (TLC) are explained as two common techniques. Key points covered include how to read a chromatogram, calculate Rf values to identify unknown substances, and factors that affect separation. The document provides instructions for homework on chromatography questions and calculations.
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Unit 2
Learning aim C: Undertake
chromatographic techniques to identify components in mixtures
LEARNING OBJECTIVES: You LO1: MUST demonstrate how to read a chromatogram. LO2: SHOULD accurately calculate Rf values on a chromatogram. LO3: COULD describe the solubility of a substance using the rF values and the correct scientific terminology. You are a newly appointed technical assistant at a large chemical plant, Chemcalequip. As part of your induction period and to progress in your role, you have to demonstrate skills in a range of practical You need to demonstrate that you can carry out different chromatographic techniques to separate and identify components in mixtures. In a report of the techniques you have used, you will need to explain the techniques, analyse and You will need to present evidence of your practical skills analysis and evaluation in a report. Criteria covered by this task: To achieve the criteria you must show that you are able to:
C.D3 Evaluate the chromatographic techniques
used in relation to outcomes and suggest improvements.
C.M3 Analyse own chromatograms and relate the
factors that affect the separation of mixtures to the quality of results obtained.
C.P5 Correctly use chromatographic techniques to
produce chromatograms.
C.P6 Explain the use of chromatographic techniques
to separate mixtures. How chromatography works In all types of chromatography, two phases are present: a stationary phase, which does not move, and a mobile phase, which flows over the stationary phase.
The mobile phase is a liquid or gas that
acts as a solvent. The substance to be tested is dissolved 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 and ethanoic acid. Dynamic equilibrium The mobile phase contains the dissolved substances of interest. It is passed over the stationary phase. As the mobile phase interacts with the stationary phase, the phases exchange particles at the same rate. A dynamic equilibrium is established.
The position of the equilibrium
determines how far the substance travels through the stationary phase. The more particles that stay in the mobile phase, the further the dissolved substance will travel. Paper chromatography 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. This technique can be used to identify additives in food. Which ink do you think? Rf values Calculating Rf values Reference material Scientists can compare the Rf value of an unknown amino acid Rf value substance to Rf values of alanine 0.38 known compounds of that serine 0.27 type. leucine 0.75 The Rf values of compounds such as amino acids are lysine 0.14 published in data books and valine 0.61 can be looked up. The amino phenylalanine 0.68 acids present can be identified by comparison of glysine 0.26 the Rf values obtained and cysteine 0.40 those in the data book. Thin layer chromatography Remember that all chromatography involves a stationary phase and a mobile phase. glass plate In thin layer chromatography (TLC) the stationary phase is a layer of silica gel fixed onto a plate. The plate may be glass, foil or plastic, for example.
The mobile phase is a solvent
that travels up the plate, silica gel carrying the substances.
In paper chromatography, what are
the stationary and mobile phases? How does TLC work? TLC uses the same basic principle as paper chromatography: capillary action draws the solvent up the plate.
A dry sample of the substance of interest is placed in the
silica gel. Then, as the solvent moves up the gel, it dissolves the sample and carries it up the plate.
The distance each substance travels depends on its affinity
(attraction) with the gel. The weaker the attraction between the substance and the gel, the further the substance will travel.
Affinity determines the position of the dynamic equilibrium
established between the mobile and stationary phases. Affinity depends on the bonding of the solvent, substance and gel. TLC or paper chromatography Thin layer chromatography has a number of advantages over paper chromatography.
1. The glass plate is rigid, not flexible like paper, so it is easy
to control.
2. After separation, the substances in the mixture can be
recovered. The silica gel holding the separated substance is scraped off the glass plate and added to a solvent. The substance will dissolve and the silica gel can easily be removed by filtration.
The glass plates can be re-coated with
silica gel and used over and over again. UV and locating agents 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. This usually works well for organic compounds.
An alternative method is to use a chemical locating agent
a chemical that reacts with the substance to form a coloured compound.
For example, when ninhydrin is exposed to an organic
compound, it stains it purple-brown. Homework Complete the questions on chromatography and calculating Rf values.
