Objective
Redo the standard curve for the Malachite green absorption and measure the fluorescence curve for Rhodamine B.
Description
- Measure UV-Vis spectrum of different malachite solutions.
- Create 2 new solutions using available stock solutions.
- Plot the data and determine extinction coefficients.
Procedure for fluorometer
- Fluorescence spectrometry is also a non-damaging technique meaning that that samples can be returned to their containers after testing.
- After cleaning the cuvette (this is done the same way as for the UV-Vis), add the sample to fill about 75% full.
- Insert the cuvette into the fluorometer.
- After saving as a .txt file to be exported, remove the cuvette from the fluorometer and return the sample to the container.
Data
- Stock solutions of Malachite green
- .309 ppm
- 1.0 ppm
- 1.5 ppm
- 5.917 ppm
- Newly created stock solutions
- 6.7 ppm (3.160g of 106 ppm Malachite green stock, water added to a total mass of 50.008g)
- 8.8 ppm (3.8277g of 106 ppm Malachite green stock, water added to a total mass of 45.659g)
The new standard curve for Malachite green
- Stock solutions for the Rhodamine B
- 66.4 ppb
- 164 ppb
- 287 ppb
- 332 ppb
- 746 ppb
- Absorption vs wavelength of Rhodamine B
- Areas under the curve for each wavelength were calculated and graphed vs the relative concentration for Rhodamine B:
Notes
- Parameters Used:
- Start(nm): 565 End(nm): 700 Excitation(nm): 555
- Ex slit(nm): 5 Em slit(nm): 50 Scan Speed(nm/min): 200
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