User:Zachary I. Mendel/Notebook/Zacks Notebook/2013/10/16

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We are going to test the activity of our HRP-NPs today for the catalytic conversion of luminol


  1. Add experimental record here. Include what, how, and why...


Stock Solution

  1. Buffer
    1. 0.6140g Tris in 1L, pH set to 8 with HCl ---> 5.1mM
  2. Luminol
    1. Dissolve 12.9mg luminol in 300uL of DMSO
    2. Add to 50mL buffer ---> 1.46mM
  3. H2O2
    1. 312uL 30% H2O2 into 100mL buffer ---> Should be 45mM
    2. Check absorption at 250 source (ε(250) = 16.69 M-1cm-1). A = 0.7392. [H2O2] = 44.29mM


Absorbance spectra of the catalytic activity of HRP-AuNPs

- We measured the absorbance of the 1/20 luminol to see if it still absorbed. We found the absorbance peak to be less than 1.00, specifically around 0.300

Graph showing the absorbance spectra for citrate AuNP and HRP AuNP

HRP and Citrate AuNP.png

Making the BSA AuNP solution

-Add 1mL of the 2.54mM gold (HAuCl4) solution to a 10mL volumetric flask

-Add an appropriate amount of BSA solution so that the final concentration of gold is 90X that of BSA

   - Calculations = ((0.00254 moles/L Gold)* (1L/1000ml)* (1ml)) / 90 = moles BSA = 2.82*10^-8 moles BSA
     [BSA] = 15.6μM
        - (2.82*10^-8 moles) * (1L/15.6*10^-6 moles) = 0.00180L or 1.8mL BSA to add to solution

Add deionized water up to 10mL

Transfer solution to a test tube and cap with aluminum foil

Heat in oven at 80C for 3 hours