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chromatography
a scientific method of finding what separate substances are in a mixture by making it flow, as a liquid or gas, through a material such as paper which different substances pass through at different speeds
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retention time
the time taken for a substance to reach the detector at the end of a gas chromatography column
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mass spectroscopy
an analytical technique that involves breaking molecules into chraged fragments and measuring their mass/charge ratio . Also know as mass spectrometry
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paper chromatography is used to
analyse coloured substances, such as the coloured pigments in plants and artificial colours used as food additives
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why does paper chromatography work
because some of the coloured substances are better at dissolving in the liquid than they are at bonding with the paper, so they travel further up the paper.
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Chromatography is used by scientists to
- Analyze – examine a mixture, its components, and their relations to one another
- Identify – determine the identity of a mixture or components based on known components
- Purify – separate components in order to isolate one of interest for further study
- Quantify – determine the amount of the a mixture and/or the components present in the sample
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Real-life examples of uses for chromatography :
- Pharmaceutical Company – determine amount of each chemical found in new product
- Hospital – detect blood or alcohol levels in a patient’s blood stream
- Law Enforcement – to compare a sample found at a crime scene to samples from suspects
- Environmental Agency – determine the level of pollutants in the water supply
- Manufacturing Plant – to purify a chemical needed to make a product
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Chromatography separates the components of a mixture by their
distinctive attraction to the mobile phase and the stationary phase.
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explain how the mobile phase and stationary phase are interlinked
- Compound is placed on stationary phase
- Mobile phase passes through the stationary phase
- Mobile phase solubilizes the components
- Mobile phase carries the individual components a certain distance through the stationary phase, depending on their attraction to both of the phases
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in paper chromatography what is the stationary phase
solvent (such as water)
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in paper chromatography what is the mobile pahse
paper strip
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how can artificial colourings added to foods be analysed using paper chromatography
a sample of the colour from the food is placed on a piece of chromatography paper along with samples of known dyes , then a solvent is added. the solvent soaks up the paper taking the dyes with it . The more soluble the colouring is , the further up the paper it travels . Different colourings move different distances up the paper
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The more soluble the colouring is
the further up the paper it travels
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why do we hang chromatography paper in water during paper chromatography
because the water is the solvent and travels up the paper taking the substance with it
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why must the solvent be below the spots during paper chromatography
so the substances don't dissolve in the water
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why was the line drawn in pencil
so the substances dont dissolve in the water
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why must you wait until the water is near the top of the paper before taking it out to dry
to allow the dyes to seperate completely
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why do the spots travel at different distances
because the dyes have different solubilities in the solvent (mobile phase) and they bond/stick to the paper (stationary phase) at varying degrees .
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a very common instrumental method is
gas chromatography
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gas chromatography can
seperate out a mixture of compounds and help you identify the substances present
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what is Rf value
- Distance moved by spot
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Distance moved by solvent
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label the diagram
- a = cylinder of carrier gas with valve to control the flow
- b = sample injector
- c = collector in which components are condensed
- d = detector
- e = chromoatogram
- f = column in temperature controlled oven
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what is the purpose of A
it contians inert gas that is the mobile phase
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what is the purpose of f
it is the column paccked with solid material , it is the stationary phase
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what is the purpose of D
it is called a chromatogram . The number of peaks tells us the number of compounds present in the mixture . The retention time varies with the different components and is used to identify the substance . The relative are under each peak tells us the relative quantity of each component
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how can substances be seperated by gas chromatography
the sample is injected into the machine and is vapourised . The sample passes through a long column packed with a solid that is wound into a coil (stationary phase) . An inert gas (unreactive) (mobile phase) such as nitrogen , is passed through the column to move the sample through . Different substances in the mixture travel through the column at different speeds as they have different affinities for the mobile and stationary phase . This means that they reach the end of the column at different times and so are sepearted .
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the greater the affinity the substance has for the mobile phase the
quicker the retention time
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the greater the affinity the substance has for the stationary phase the
slower the retention time
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a mass spectroscopy can
identify tiny amounts of a substance quickly and accurately it does this by measuring the mass of the particles in the substance
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in mass spec the molecules
lose an electron to form a molecular ion . the Mr of the substance = the mass of the molecular ion . often the molecular ion breaks apart and other lighter fragments are detected aswell .
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a mass spec is often attatched
to a gas chromatography machine . after a smaple has been seperated by chromatography , the mass spectrum of each compound can be recorded allowing it to be identified
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which peak gives the Mr and what is the substance
the peak furthest to the right is the molecular ion peak and gives the Mr of the substance - this means the Mr of this substance is 204 , this substance is mercury
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