spectrophotometry

spectrophotometry - set transmittance to 100% record %...

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1 SPECTROPHOTOMETRY Introduction • Electromagnetic radiation gamma rays x-rays UV(visible light = 380 nm to 750 nm) infrared TV and radio waves • the smaller the wavelength, the greater the energy 2 Visible Light • violet 380 nm •b lue • green •ye l low •o range • red 750 nm 3 Parts of the Spec 20 • Wavelength control knob
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4 Visible Portion of the Electromagnetic Spectrum • Set wavelength to 620 nm • Record wavelenght ranges for: red, orange, yellow, green, blue, violet light 5 Using the Spec 20 •wa rm up Spec 20 •make • blank = 1ml water + 5 ml indicator •samp le = 1ml protein solution + 5 ml indicator (protein solution = protein + water)
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Unformatted text preview: set transmittance to 100% record % transmittance and absorbance *check lever of some machines 7 Development of a Standard Curve Tube # Water Protein soln. Indicator *1 0 1 ml 5 ml *Already preparedsample tube Remove 1 ml = 2 ml, mix 1 ml 5 Remove 1 ml = 2 ml, mix 1 ml 4 Remove 1 ml = 2 ml, mix 1 ml 3 Remove 1 ml = 2 ml, mix 1 ml 1 ml 2 Protein soln. Water Tube # dispose 8 Development of a Standard Curve Add indicator at the end after tubes have 2ml of mixture 5 ml 2 ml 5 5 ml 2 ml 4 5 ml 2 ml 3 5 ml 2 ml 2 indicator Water + protein soln Tube # 9 Results Plot protein concentration in mg/ml vs. absorbance on graph paper measure absorbance of unknown and determine its protein concentration in mg/ml (extrapolate from graph) 10 Graph X-axis (independent variable) protein concentration in mg/ml Y-axis (dependent variable) Absorbance...
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spectrophotometry - set transmittance to 100% record %...

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