A new model for Overhauser enhanced nuclear magnetic resonance using nitroxide radicals

Abstract

Nitroxide free radicals are the most commonly used source for dynamic nuclear polarization (DNP) enhanced nuclear magnetic resonance (NMR) experiments and are also exclusively employed as spin labels for electron spin resonance (ESR) spectroscopy of diamagnetic molecules and materials. Nitroxide free radicals have been shown to have strong dipolar coupling to (1)H in water, and thus result in large DNP enhancement of (1)H NMR signal via the well known Overhauser effect. The fundamental parameter in a DNP experiment is the coupling factor, since it ultimately determines the maximum NMR signal enhancements which can be achieved. Despite their widespread use, measurements of the coupling factor of nitroxide free radicals have been inconsistent, and current models have failed to successfully explain our experimental data. We found that the inconsistency in determining the coupling factor arises from not taking into account the characteristics of the ESR transitions, which are split into three (or two) lines due to the hyperfine coupling of the electron to the (14)N nuclei (or (15)N) of the nitric oxide radical. Both intermolecular Heisenberg spin exchange interactions as well as intramolecular nitrogen nuclear spin relaxation mix the three (or two) ESR transitions. However, neither effect has been taken into account in any experimental studies on utilizing or quantifying the Overhauser driven DNP effects. The expected effect of Heisenberg spin exchange on Overhauser enhancements has already been theoretically predicted and observed by Bates and Drozdoski [J. Chem. Phys. 67, 4038 (1977)]. Here, we present a new model for quantifying Overhauser enhancements through nitroxide free radicals that includes both effects on mixing the ESR hyperfine states. This model predicts the maximum saturation factor to be considerably higher by the effect of nitrogen nuclear spin relaxation. Because intramolecular nitrogen spin relaxation is independent of the nitroxide concentration, this effect is still significant at low radical concentrations where electron spin exchange is negligible. This implies that the only correct way to determine the coupling factor of nitroxide free radicals is to measure the maximum enhancement at different concentrations and extrapolate the results to infinite concentration. We verify our model with a series of DNP experimental studies on (1)H NMR signal enhancement of water by means of (14)N as well as (15)N isotope enriched nitroxide radicals.

Studies of Dynamic Nuclear Polarization with Nitroxides in Aqueous Solution

“…This saturation factor is higher than the expected one when saturating only one of the three hyperfine lines (s " 1/3). In agreement with the proposal of other authors [13] (although our reported x are substantially different), the large saturation factors can be interpreted in terms of an electronelectron double resonance effect which is caused either by the very fast nuclear spinlattice relaxation of the strongly coupled nitrogen or by intermolecular Heisenberg spin exchange. Both mechanisms compete with the internal electron T 1 relaxation and cause indirect saturation of the adjacent hyperfine lines [14,15].…”

First DNP Results from a Liquid Water-TEMPOL Sample at 400 MHz and 260 GHz

“…(1), DNP measurements should be performed with changing radical concentration and different hyperfine couplings by replacing 14 N with 15 N [15]. Therefore no reliable direct estimates for the coupling factor x can be made in this paper.…”

Saturation factor of nitroxide radicals in liquid DNP by pulsed ELDOR experiments

“…[8][9][10][11][12] Due to the strong hyperfine coupling between the electron and the nitrogen nucleus, which splits the EPR spectrum into two ( 15 N) or three ( 14 N) separate lines, the saturation factor, defined as s eff = (S B À hS z i)/S B with S B the Boltzmann polarization of the electron spin, cannot be simply extracted from the saturation behaviour of the pumped line according to the Bloch equation, as formerly suggested for trityl radicals. 13 Moreover, the expectation value of the electron spin polarization hS z i depends on the population of all energy levels involved.…”

“…the ratio of the gyromagnetic constants of the electron spin g s and the target nucleus g I , the coupling factor x, the leakage factor f and the effective saturation factor s eff of the EPR line: 6 e = 1 À s eff fx|g s |/g I (1) In recent studies, nitroxide radicals have been favoured as polarizing agents for DNP since they are soluble in water, well compatible with biological systems, non-toxic and have been found to account for large DNP enhancements at magnetic fields up to 9 T. 7,8 However, the determination of the saturation factor for this class of polarizers has emerged as one of the major difficulties in rationalizing the observed DNP enhancements in terms of the Overhauser equation (1). [8][9][10][11][12] Due to the strong hyperfine coupling between the electron and the nitrogen nucleus, which splits the EPR spectrum into two ( 15 N) or three ( 14 N) separate lines, the saturation factor, defined as s eff = (S B À hS z i)/S B with S B the Boltzmann polarization of the electron spin, cannot be simply extracted from the saturation behaviour of the pumped line according to the Bloch equation, as formerly suggested for trityl radicals. 13 Moreover, the expectation value of the electron spin polarization hS z i depends on the population of all energy levels involved.…”

“…13 Moreover, the expectation value of the electron spin polarization hS z i depends on the population of all energy levels involved. In recent years, this issue has been debated in the literature and several theoretical models were proposed to calculate the saturation factor for nitroxides [9][10][11] but to date no direct experimental approach has been provided.…”