Exchangeable proton signals

NOE connectivities provided the shifts of non-exchangeable proton signals of these His imidazole rings. Their shifts are summarized in Table 2. The similarity in the shifts between metMb(OH ) and diamagnetic Mbs, i.e. Mb(CO) and oxy-Mb (MblO )), demonstrates that paramagnetic contribution to these resonances is quite small. Semiquantitative analysis of 6pc, due to... 

Effects of salt concentration on high-frequency shifted exchangeable proton signals... 

Fig. 3. Rapid Mo(V) EPR signals obtained on reducing xanthine oxidase at pH 10 with 15 moles of xanthine for 1 min. at about 20 °. The upper four spectra are at 9.1 GHz and the lower four at 34.4 GHz. a, a, c, 8 refer to H2O as solvent and b, b, d, d to D2O. a, b, c, d are computer simulations of the experimental spectra, a, b, c, d, respectively. The interpretation is that two species, each having exchangeable protons which interact with Mo(V), are responsible for the signals. For one of these (dotted complex type II) there are two equivalent interacting protons and for the other (dashed complex type I), two non-equivalent protons. These species are believed to correspond to two different complexes of reduced xanthine oxidase with xanthine. (Reproduced from ref. 78 see also Table 2 for the parameters of the signals.)...

To continue the investigation, carbon detected proton T relaxation data were also collected and were used to calculate proton T relaxation times. Similarly, 19F T measurements were also made. The calculated relaxation values are shown above each peak of interest in Fig. 10.25. A substantial difference is evident in the proton T relaxation times across the API peaks in both carbon spectra. Due to spin diffusion, the protons can exchange their signals with each other even when separated by as much as tens of nanometers. Since a potential API-excipient interaction would act on the molecular scale, spin diffusion occurs between the API and excipient molecules, and the protons therefore show a single, uniform relaxation time regardless of whether they are on the API or the excipients. On the other hand, in the case of a physical mixture, the molecules of API and excipients are well separated spatially, and so no bulk spin diffusion can occur. Two unique proton relaxation rates are then expected, one for the API and another for the excipients. This is evident in the carbon spectrum of the physical mixture shown on the bottom of Fig. 10.25. Comparing this reference to the relaxation data for the formulation, it is readily apparent that the formulation exhibits essentially one proton T1 relaxation time across the carbon spectrum. This therefore demonstrates that there is indeed an interaction between the drug substance and the excipients in the formulation. 

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