User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/09

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(Chemiluminescence of Luminol)
(Chemiluminescence of Luminol)
Line 33: Line 33:
<math>\frac{53.148 g}{10}</math> = 5.3148 g of luminol in 1000 mL of water (30 mM)
<math>\frac{53.148 g}{10}</math> = 5.3148 g of luminol in 1000 mL of water (30 mM)
x = <math>\frac{2 mL * 5.3148 g}{1000}</math> = .0106296 g of luminol in 2 mL of water (30 mM)
==Fluorescence Spectrometer Graph==
==Fluorescence Spectrometer Graph==

Revision as of 22:49, 25 October 2012

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Atomic Absorption Spectroscopy of Au/BSA

  • A rack of Au/BSA solutions was recovered and used for the assigned Atomic Absorption Spectroscopy (AAS). AAS was used to strip the molecule to its elemental state.
  • The entire solution set were observed to lack fibers even at high mole ratios where formation of fibers is typical. The date of which the solutions were made is unknown.
  • The equipment used for AAS is by the make of Shimadzu AA-6200 Atomic Absorption Flame Emission.
  • The sample cell used for this procedure is the ignited flame.
  • A gold hollow cathode tube was selected for the procedure. The wavelength of the lamp was 242.8 nm.
  • Two blanks were used; water and HCl. Water was chosen since it is part of the ternary mixture. The standards provided by Dr. Miller were made with HCl. In addition, HCl ionizes better and keeps the metal in solution. The standards were 5, 8, 10, 15, 20, 25, 30, 40 ppm Au which appeared as clear and colorless liquid solutions.
  • Measurement was done by inserting a transparent, plastic tube attached to the equipment into the solution.
  • Each blank, water and HCl, was measured and followed by autozero. Next, standards 1, 2, and 3 were measured.
  • The flame is blue green in its normal state. Having run the blanks, the flame remained blue green during the re-measurement of each blank.
  • During the measurement of the standards, the flame was observed to turn orange. The color orange is indicative of the ions present in the solution.
  • The other standards and samples were run. The calibration curve is on Puja's notebook.

Chemiluminescence of Luminol

  • The reaction of 3-aminophthalhydrazide (luminol) and hydrogen peroxide is catalyzed by the addition of horseradish peroxidase (HRP) producing 3-aminophthalate.
  • The reagents originated from the HRP Assay of aminoantipyrine (AAP) laboratory period 10/02/2012. For this procedure, 4-iodophenol, hydrogen peroxide, HRP, and water will be used. The AAP will be replaced in the reaction with luminol.
  • It was decided to prepare 10 mM of luminol since this will be further diluted down to 3 mM upon the addition of water in the cuvette.
  • The following calculations were made for luminol (FW 177.16):

\frac{177.16 g}{10} = 17. 716 g of luminol in 1000 mL of water (.1M)

17.716 g × 3 = 53.148 g of luminol in 1000 mL of water (.3 M = 300 mM)

\frac{53.148 g}{10} = 5.3148 g of luminol in 1000 mL of water (30 mM)

x = \frac{2 mL * 5.3148 g}{1000} = .0106296 g of luminol in 2 mL of water (30 mM)

Fluorescence Spectrometer Graph


No luminescence was observed for any runs.

The concentration of reagents are shown below the table:

sample 4-iodophenol luminol H2O2 H2O
130 μL900 μL176 μL1.8 mL

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