Journal of Physical Chemistry A, Vol.102, No.45, 8749-8758, 1998
Fast time-resolved magnetic optical rotatory dispersion measurements. 2. Confirmation of Mueller analysis and application to myoglobin ligand photolysis
Experimental verifications are presented of expressions derived in the preceding paper that describe a simple and sensitive near-null technique for nanosecond time-resolved magnetic optical rotatory dispersion (TRMORD) spectroscopy. Expressions verified include those describing artifacts associated with strain, imperfect Faraday compensation, and photoselection-induced orientation of the sample. The accuracy of MORD data obtained with this method is demonstrated by comparison with MORD curves calculated from the Kramers-Kronig transform of independently measured magnetic circular dichroism (MCD) spectra. The separation of natural and magnetic optical rotations for chiral biological macromolecules and potential artifacts associated with this separation are discussed. This work confirms the prediction in paper 1 in this series that the MORD technique is less sensitive to linear birefringence (arising from Strained optics or photoselected samples, for instance) than the related near-null MCD technique previously developed in this lab. Possible problems associated with imperfect Faraday compensation are discussed, as is the prediction in the preceding paper of a coupling between photoselection-induced linear dichroism of the sample and Faraday rotation of the solvent. Photolysis measurements on carbonmonoxymyoglobin demonstrate that an excitation geometry in which the sample is probed along the unique axis of excitation minimizes the latter effect, improving accuracy in MORD measurements on photoselected samples. The TRMORD technique is applied to the ligand rebinding reaction of photolyzed carbonmonoxymyoglobin in Soret-region spectral measurements that demonstrate the ability of this technique to measure multichannel time-resolved magnetooptical data with a precision suitable for global kinetic analysis.