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A fast freezing technique using liquid propane was used to obtain frozen solutions of methemoglobin at pH 7. With this method it was possible to eliminate largely the presence of a low spin component which is usually found in slowly frozen solutions but not present in the sample at room temperature. The magnetic susceptibility and Mossbauer spectra of Hb(H 2 O) in the temperature range 4.2 K ≦ 7 ≦ 250 K have been measured. The data of the high spin compound of Hb (H 2 O) were evaluated with a Hamiltonian containing the Coulomb repulsion of the five 3d-electrons of the Fe 3+  ion, a crystal electric field of C 2v  symmetry and the spin orbit coupling. The term describing the crystal electric field depends on five energy parameters ε 1  , ε 2 , ε 3 , D, and E which are determined by least squares fits to the experimental data. The most relevant parameters ε 2  and ε 3  which equal the energies of the antibonding single 3d-electron orbitals 3d z  2  and 3d x  2 -y 2  with respect to the 3d sy  orbital are compared with earlier results of these energies ε 2  and ε 3  of the high spin compound of Mb(H 2 O). From this comparison conclusions regarding the different spatial ar­rangements of Fe 3+  in Hb(H 2 O) and Mb(H 2 O) are drawn.

The Active Center of Methemoglobin Hb (H 2 0 ) Investigated by Mössbauer and Susceptibility Experiments