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A-level Applied Science/Finding out about substances/Colorimetry
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Uses of colorimetry
Colorimetry is used in both biology and chemistry and in other sorts of places such as in industries, color printing, textile manufacturing, paint manufacturing and in food industries (including the chocolate industry). Colorimetry is also used in aspirin.
Colorimetry can detect the smallest colour difference that the human eye cannot pick up.
Under the action of chemical agents, samples develop a specific colour that shows the concentration of the substance being tested.
Colorimetry is just one of the types of photometric analysis techniques i.e. it is a way of measuring light.
Below are most possible colorimetric tests:
Colorimetry can be used to find out the concentration of any coloured substance.
Applications of colorimetry
Food & Beverage Quality Control
Alpha Amylase Activity
Milk Quality
Miscellaneous Quality Tests
Mineral Oils & Fuels
Anti-Icing Additive in Aviation Fuels
Marked Oils
Carbonisable Substances
Lead content
Medical & Clinical Tests
Alpha Amylase in Blood or Urine
Bilirubin
Cholesterol
Cholinesterase, Activity in Blood
Haemoglobin Content of Blood cells
Iron in Serum
Lactate Dehydrogenase
Lead in Urine/Feaces
pH Value of Blood/Urine
Phenolsulphophhtalein (Phenol Red) Excretion Test
Phenylpyruvic Acid
Phosphatase in Blood
Phosphorus, Inorganic, in Blood
Proteins
Salicylate In Blood
Sugar/Glucose in Blood
Sulphobromophthalein Retention
Sulphonamides in Blood/Urine
Trichloroaccetic Acid
Urea in Blood
Uric Acid in Serum
Air Monitoring Tests
Chromium
Lead
Pipino
Method
Colorimetry measurements are made by using a light which passes through a colour filter. The light then passes through a little box (cuvette) with the actual chemical substance. The light leaving the actual sample should be less than the light that actually entered the compound. The loss of light always reflects the concentration of the compound.
Colorimetry can only be done to measurements which are within the visible region of the electromagnetic spectrum, which is 380 – 780 nm. The main general factors which affect the amount of light absorbed by the sample are the wavelength of the light and the colour of the solution.
When manufactures are using a colorimeter, they have a choice of which wavelength they use. The options are:
Gelatin filters
Interference filters
Grating Monochromators
Prisms
Sample preparation
Weighed into the crucible, the cotton thread is attached to the wire, and the crucible is put into the vessel. This applies to powdery substances
Colorimetric Measurement of Iron Concentrations
By making several solutions of iron (II) sulfate with known concentrations, the absorbance of each can be determined and a calibration curve can be constructed. Given a solution of unknown concentration, its concentration can be determined from its absorbance.
You will be given iron (II) sulfate solution of known concentration.
Equipment
Volumetric flasks, 100cm³
Colorimeter
graduated Pipette
Pipette filler
cuvette
Reagents
Iron (II) sulfate (0.100 mol dm-3)
Precautions
Wear eye protection, protective gloves and laboratory coat at all times.
Make sure you wear shoes.
Make sure you dont put the substance in your eyes.
Procedure
Use the colorimeter to measure the absorbance of the water sample and the standard solutions across a range of wavelengths. Choose a suitable wavelength for the rest of the experiments.
As a group, decide on how you will need to dilute the standard solutions. You will need to make a series of dilutions which will have absorbances similar to the water sample.
Use a colorimeter to determine the absorbance, at the appropriate wavelength of every diluted standard solution made.
Results
Plot a graph of absorbance against wavelength for the unknown sample and the standard solutions.
Explain your choice of wavelength.
Plot a graph of absorbance against concentration for the diluted standard solutions.
Calculate the concentration of the unknown solution.
Comment on the shape on the graph: According to the Beer Lambert Law it should be a straight line passing through the origin (i.e. directly proportional).
Risk assessment
Experiment or Activity: Colorimetry
Material used or Procedure:Hazard
(the harm it can cause):Risk
(the chance of harm):Reducing the risk
(What can be done to make it safer):Emergency Procedures (What to do if something goes wrong, e.g. First Aid)
Nitric acid
Phosphoric acid
Potassium iodate
Potassium mangonate
All of these are harmful to skin. If in contact with skin they are an irritant. Also can cause burns to skin if left in contact too long.
Also can get in eyes and cause blindness
Wear protective gloves when handling acid. Wear protective goggles at all times when conducting experiment.
If in contact with skin run under cold water immediately. If acid gets in eyes rinse and wash out eyes.
Observations and measurements
once you have carried out they experiment your results should show a clear pattern. As the concentration of the solution increases so should the absorbance recorded, meaning they are proportional to one another.
Results, calculation and evaluation
Percentage %Unknown (A)0.1 M (A)
100% 0.13 0.29
75% 0.10 0.24
50% 0.08 0.15
25% 0.06 0.11
Evaluation of our colorimetry experiment:
The good thing about our experiment is that we got good results and it compared well against the rest of the class because there percentages as seen of the graph were around the same mark as ours was. A bad thing about our colorimetry experiment is that the colorimeter was playing up and sometimes gave us the wrong readings and this set us back a bit because we had to change the colorimeter meaning again we could not repeat.
Scientific principles behind colorimetry
Colorimetry is the science that describes colours in numbers. It is used in chemistry, and in industries such as colour printing, textile manufacturing, and paint manufacturing.
A colorimeter measures the intensity of light shining through a coloured solution compared to the intensity of light passing into the solution. A detector measures the transmittance (T) (% of light passing through) of the solution. This is mathematically converted to absorbance (A = #log10T). The absorbance is directly proportional to the concentration (Beer#Lambert law). The colorimetry of sulfuric acid is a blue colour and will give an absorption of 0.42 when doing serial dilution.