Spectrophotometric Analysis of Kmno4 Solution

After the serial dilutions of the red and blue dyes were taken, the molarity and absorbance for both dyes were calculated. Using the MiVi = MfVf equation, the concentrations for each value of the red and blue dye were separately calculated. Calculating absorbances calls for setting the correct wavelengths of light for each dye. In this case, the 470 nm wavelength for red dye and the 635 nm wavelength for blue dye was needed to find the maximum absorbances. The absorbance was found by blanking the colorimeter and entering the concentrations. After both values of the absorbances and concentrations were found, the values were then graphed in order to obtain the equation of the relationship between absorbance and concentration.

From this graph and chart we can see that the higher the concentration the higher the absorbance, all the different concentrations were tested at the same wavelength (625nm). Also we can determine our unknown substances concentration by using the absorbance we got for it. The red dot on the graph followed by the line towards the horizontal axis indicates that the concentration of fast green was 34% or 5.1x10-3.

10 microliters of the sample is then added and the assay absorption is measured at 340nm. If absorbance was above 1.5, samples were diluted.

Colorimetric assay is a process determining the concentration of a chemical element or compound in a solution

A spectrophotometer’s purpose is to use colors of the light spectrum to determine the concentration of light absorbing molecules in a solution. (p.59) In this particular lab, our mission was to determine the protein concentration and the standard curve of the unknown sample of BSA. This, by preparing five dilutions of the unknown solution of BSA together with other known concentrations, and then experimenting by observing how the concentrations were passed through the spectrophotometer. The outcome resolved in the absorption levels being decreased, and this

Time) because it had a correlation closest to 1. All three orders were graphed and a linear regression was used to see which graphed order was closest to 1. The order was determined by comparing the concentration and time to the mathematical predictions made using the integrated rate laws. Analyzing each graph and finding each correlation helped determine which graph was closest to 1. The more concentrated a solution is, the higher the absorbance of that solution. This is due to Beer’s Law. The law measures the absorbance of a solution by determining how much light passes through a solution. As the concentration of a solution increases, fewer wavelengths of light are able to pass through the concentrated solution. The absorbance at 60 seconds was 0.573 (Figure 1: Table1). To calculate the concentration (molarity), the Beer’s Law equation was used, Abs = slope(m)+b. Plugging in what is known into the Beer’s Law equation resulted in 0.573 = 3.172e+004 + 0, where the concentration is determined by M = 0.573-0/ 3.172e+004. So, the concentration at 60 seconds using the equation (M = 0.573-0 / 3.172e+004) was 1.824e-5 M. The 1st order graph resulted in k=0.006152 (Figure 1: Graph 1). Other groups also resulted in their decolorization of CV to be the 1st rate

Scientists use an instrument called a spectrometer to quantitatively determine the amount of light absorbed by a solution. The primary inner parts of a typical spectrometer are described below. The spectrometer has a light source that emits white light containing a vast mixture of different wavelengths of electromagnetic radiation. The wavelength of interest is then selected using a monochromator (“mono” meaning one and “chromate” meaning color) and an additional exit slit. The separation of white light into different colors (wavelengths) is known as diffraction. The selected light then reaches the sample and depending on how the light interacts with the chemical compound of interest, some of the light is absorbed and some passes straight through. By comparing the amount of light entering the sample (P0) with the amount of light reaching the detector (P), the spectrometer is able to tell how much light is absorbed by the sample.

The Beers Law calibration experiment used many concentrations of crystal violet solutions. Each of these solutions were test and analyzed in order to determine the absorbance of each concentration The results were than graphed and produced a slope of 1.00E05 with an intercept of -2.21E-02.

Pour 50 mL of distilled water into a 100 mL beaker, and then add the unknown substance into it. Mix thoroughly to create the aqueous solution. Now fill a new cuvette with this new solution and place into the SpectroVis Plus device; after wiping the outside with a Kimwipe as usual. Be sure to take note of the absorbance when wavelength is at its maximum. Afterwards, Be sure to take all solutions containing Iron(III) and pour them into the container specified for hazardous wastes. ​

The wavelength of the light should be a different color from the solution’s color. This is because if the color of the wavelength of the light is the same color as the solution’s color, then the color would not be absorbed. The only way that a solution can absorb a color is if the color of the light’s wavelength is its complementary (opposite) color. The color of the light chosen for this experiment was blue because the wavelength was set to 430nm, which corresponds to blue's wavelength. The color of the FeSCN2+ complex ion is blood-red.

When you use a spectrophotometer you should not set the wavelength of light to be the same color of the solution. This is because if you set the wavelength to be the same color of the solution then no light will be absorbed. The reason why no light will be absorbed is because the color you see is the wavelength of light that is being reflected so you must set the wavelength to be the complementary color. The wavelength of light that was chosen for the lab was 450nm which coincides with a very dark blue color. The reason why choosing a dark blue makes the most sense for this experiment is because the color of the FeSCN2+ ion was blood red.

A = Absorbance difference  = Molar extinction coefficient C = Concentration L = Path length

A spectrophotometer is an instrument which measures the amount of light of a specified wavelength which passes through a medium. This instrument is usually used for the measurement of reflectance of solutions. Light is separate into different wavelengths and is being passed through the sample solution. The sample solution will have its own wavelength and will absorb a certain amount of light. The higher the molecular concentration, the higher the absorbance value.

8) Steps 1 - 8 were repeated using the wavelengths of 360 nm to 900

concentration, record the absorbance readings at a fixed wavelength, and plot the absorbance vs. concentration data. The wavelength of 520 nm was selected for experiment Part