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Four Steps from Raw Data to Final Answer

Step One: Start with Recorded Data
(from Data Collection or Saved Files)

Gather multi-wavelength data to "record" the changes in a sample.

The driving force of the change in the sample can be:

  • time - kinetic experiments
  • temperature - denaturation experiments
  • pH - titration experiments (scans as a function of time or temperature or pH)

Optical measurements can be:

  • Absorbance
  • Fluorescence
  • Circular Dichroism
  • ORD
  • FTIR

Step Two: Confirm Number of Species

Apply SVD to the 3D data set.

SVD is Singular Value Decomposition
(a form of factor analysis).

  • Indicates the number of spectrally distinct species involved in the recorded change
  • Greatly reduces the complexity of the fitting
  • Dramatic noise reduction


Step Three: Select the Chemical Relationship of Species

Fit the data to an appropriate equation.

If the data are kinetic, the Olis program offers many kinetic fit equations derived from a wide variety of mechanisms. Equilibrium data involving temperature or pH effects can also be fitted with different mechanisms.


Step Four: Final Answer is Spectra and Kinetics of Species

After fitting, compute spectra for the reacting species.

If the mechanism chosen results in the creation of credible spectra for the reacting species, the correctness of the choice is reinforced; if credible spectra are not produced, that mechanism cannot be correct.


 

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