Spectrophotometry Electromagnetic Radiation = Light What is Light?

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Presentation transcript:

Spectrophotometry Electromagnetic Radiation = Light What is Light? Visible light is a particular kind of electromagnetic radiation X-rays, UV, Infrared, Microwaves, and Radio waves are all light forms Light is a form of Energy All light travels in waves at “the speed of light” = c = 3 x 108 m/s 2

Characteristics of Light Wavelength = distance between two peaks in a wave l (lambda) is the symbol Meters = m is the unit Frequency = number of complete waves passing a given point per second n (nu) is the symbol Hertz = Hz = s-1 is the unit Amplitude = A = measure of the intensity of the wave, “brightness” The speed of light is constant: c =  x  l and n are inversely proportional If one increases, the other decreases Their product is always the speed of light = c =  x  = 3 x 108 m/s

II. Spectroscopy = using light to investigate a compound How the experiment works What color do we see? a) What we see as the color of a compound is the complementary color to what the compound absorbs b) Example: Absorbs red, we see green

Beer’s Law Absorbance tells us how much light is absorbed by the molecule B. A = elc (y = mx + b) describes the absorption of light in a solution l = the length of the cell containing the solution, usual 1 cm c = concentration in mol/L = M e = Molar extinction coefficient = constant for a given molecule at a given wavelength of light = how well the molecule absorbs light Percent T 0 10 25 50 75 100 2.0 1.0 0.6 0.3 0.1 0 Absorbance 1.0 0.5 0.0 e ≈ 18,000 for our red dye Absorbance Slope = el [L/mol∙cm][cm] 0 0.00001 0.00002 0.00003 0.00004 0.00005 Concentration (mol/L)

Today’s Experiment Make a Beer’s Law Calibration Curve using a Stock Solution Calculating the concentration of your stock solution: (Ex: 0.0788g/L) Calculations for the calibration curve samples: M1V1 = M2V2 An Excel Template is available at faculty.swosu.edu/tim.hubin in the “Shared Folder” link called “Spectrophotometry Lab Excel sheet” Sample # 1 2 3 4 5 6 mL Dye Stock Soln 0.50 1.00 1.50 2.00 2.50 3.00 mL Water 9.50 9.00 8.50 8.00 7.50 7.00 Molarity

Find the concentration of an unknown sample of the red dye We will give you an unknown amount of the stock solution Dilute it to 25 ml in a volumetric flask Calculate the absorbance (from %T) on the Spec. 20 Use your spreadsheet to calculate the concentration from the absorbance Example Calculation: %T = 15% for your diluted unknown Make 100ml of a solution of a known concentration (2.5 x 10-5 M)

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