Serial Dilution Activity Using a pipet Simulated Acid: Red = nitric acid Yellow = sulfuric acid Blue = hydrochloric acid.

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Presentation transcript:

Serial Dilution Activity Using a pipet Simulated Acid: Red = nitric acid Yellow = sulfuric acid Blue = hydrochloric acid

Spec-20 Instrument Sample holder cover Amplifier control knob Light control knob Wavelength control knob

Spec-20 Absorbance (wavelength) Absorbance 100 % Absorbance 0 % Transmittance 0 % Absorbance 100 % Transmittance Insert Photograph Spec-20 Sample-holder Meter Zero adjust and power control Wavelength scale Wavelength control Light control

Schematic representation of a spectrophotometer Detector Sample-holder Meter Zero adjust and power control Wavelength scale Wavelength control Light control Monochromator Light Source (1) Sample cell (3) (2) (4) (5) IoIo I Meter A spectrophotometer is an instrument that measures the fraction (I/I o ) of an incident beam of light (I o ) which is transmitted (I) by a sample at a particular wavelength. For a given substance, the amount of light absorbed depends on: a) the concentration; b) the cell or path length; c) the wavelength of light; and d) the solvent.

Wavelength (nm) Absorbance Absorbance of Chlorophyll Frequency (Hz) Wavelength (nm) cosmic rays gamma rays x-raysultra- violet infrared radio (microwave) radartele- vision radiopower transmission VioletBlueGreen YellowOrangeRed UV Near Infrared 400 nm 500 nm 600 nm 700 nm

Kelter, Carr, Scott,, Chemistry: A World of Choices  1999, page 480 Amount absorbed 663 nm Wavelength (nm)

Calibration Curve Concentration Absorbance 100 % Absorbance 0 % Transmittance 0 % Absorbance 100 % Transmittance out of linear range (too concentrated) x 2 (fixed wavelength) ? Dilute sample with water 50:50. Run sample, read concentration.

Dilution of Solutions Keys Dilution of Solutions