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Why Potassium dichromate is used for calibrate the control of absorbance of UV spectrophotometer?
Ritvikvenkat
Potassium dichromate is used as the primary standard for UV spectrophotometry because of its properties. It is pure, stable, has no waters of hydration, and has a high molar mass.
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∙ 9y ago
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How do you calibrate spectrophotometer?
press the bottom and there you go
How can one calibrate a UV spectrophotometer?
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A spectrometer is normally a system that detects changes in the way light passes thru a sample. These machines are utilized largely inside research labs in Universities, private companies, and professional industries. There are hundreds of different types of these machines. However all of them work the same way.Do I really need to calibrate a UV VIS spectrophotometer each time I use it?The short answer to this question is no, you don't have to calibrate each and every time you use your machine. However, making sure the spectrophotometer or spectrometer is calibrated correctly is of the utmost importance. If the machine is giving incorrect readings, then researchers and scientists will just be wasting precious time and money. Also calibrating will tell you if your light source(s) are getting old and need to be replaced. Difficulty: Super EasyInstructionsTools you'll need:UV VIS Spectrophotometer Calibration StandardsSpectrophotometer or SpectrometerCertificate of CalibrationStep 1: Turn on the spectrometer or spectrophotometer and let it warm up for at least 15 minutes. Check your manual to see if your machine has a longer or shorter warm-up period.Step 2: Choose the wavelength that you need to calibrate.Step 3: Select the appropriate calibration filter thickness. This can be found in the owner's manual in the "How to Calibrate A UV VIS Spectrophotometer" Section.Step 4: Inspect the calibration filter to ensure no dust, oils, or debris is on the optical surfaces.Step 5: Carefully load the filter into the cuvette holder. Close the lid and wait for the measurement.Step 6: Compare the results to what is specified on the manufacturers Certificate of Calibration.Step 7: If the numbers match within +/- 5%, your spectrophotometer/spectrometer is calibrated correctly.Step 8: If the numbers are not within the +/-5% tolerance, then adjustments need to be made to your machine.
How is a spectrophotometer calibrated?
Spectrophotometersare useful devices that allow you to mainly find out the concentration of molecules within a reaction mixture. This mixture could be an enzyme and its substrate, or much simpler chemicals.Most spectrophotometerswork using light from the U.V-visiblepart of the electromagnetic spectrum (150-700nm). The radiation hits the sample. Some parts of the sample molecules, called chromophores,absorb some of this radiation. A detector measures how much radiation has been absorbed by the sample.One can then use the Beer-Lambertlaw to calculate the concentration of your chemical sample. The law is given by A=EclA=Absorbance,E=Molarextinction coefficient (constant unique to different chromophores)c=concentrationl=pathlength (usually 1cm- the size of the reaction vessel). From the absorbancevalue given by our sample, and its molar extinction coefficient, one can then rearrange the equation to find c:c=A/E.lFrom this, one can work out the rates of chemical reactions etc.N.B-As a side-note,the technique is cheap, easy and reliable. Also, The 'E' above is notated as a Greek letter 'Eta' in other reference sources, just in case you read up more about the Beer-LambertLaw.
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