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Current Analytical Chemistry, 2005, 1, 3-27 3

Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy for


the Investigation of Supra- and Biomolecular Systems
Ingo Schnell*

Max-Planck-Institut für Polymerforschung, Postfach 3148, D-55021 Mainz, Germany


Abstract: After the fields of liquid- and solid-state NMR spectroscopy have developed largely independently
of each other over the past 50 years, materials of current interest call for new approaches which combine and
merge spectroscopic techniques from either side. A particular possibility to promote NMR spectroscopy into
this direction is provided by magic-angle sample spinning (MAS), which can be performed either at high
speeds in solid-state NMR or at moderate speeds in combination with high-resolution NMR. In this way, new
NMR methods and experiments have recently been developed, as will be discussed in this review.
In solid-state NMR, new and versatile experiments are based on coherent spin-states and coherent transfer
mechanisms, provided that fast MAS ensures efficient averaging of anisotropic interactions, and recoupling
schemes selectively re-introduce the interactions of interest. In high-resolution NMR, MAS can be borrowed
from solid-state NMR in order to ensure sufficient spectral resolution for samples with residual anisotropies or
heterogeneities (so-called HRMAS). In such HRMAS studies, recoupling techniques become applicable and
provide fully quantitative information on molecular structure and dynamics.
Keywords: NMR spectroscopy, Fast magic-angle spinning (MAS), High-resolution magic-angle spinning (HRMAS), Dipolar
recoupling, Double-quantum coherences, Polarisation transfer.

1. INTRODUCTION NMR spectra of such samples consist of very sharp


resonance lines which provide detailed information on the
The past 60 years have seen the tremendous development solubilised molecules. The chemical shift allows nuclei of
of nuclear magnetic resonance (NMR) from its discovery by the same isotope to be sensitively distinguished according to
P URCELL, P OUND [1] and B LOCH [2] in 1945/46 to one of their individual chemical environment, while the J-couplings
the most powerful characterisation methods with widely reflect the network of chemical bonds. These effects form the
spread applications in all areas of science. In the course of basis for high-resolution NMR spectroscopy, whose
this development, three major areas of NMR research development into a uniquely powerful characterisation
activities emerged: liquid-state (or high-resolution) NMR method was particularly driven by the ongoing quest for
spectroscopy, solid-state NMR spectroscopy, and magnetic- homogeneous, superconducting high-field magnets, the
resonance imaging (MRI). When NMR methods and introduction of pulsed Fourier-transform spectroscopy by
techniques became more and more sophisticated and ERNST in 1971 [7, 8] and the extension to multidimens-
specialised, the separation of these “principal” NMR areas ional spectra proposed by JEENER[9] and pioneered by
intensified such that they are largely considered as ERNST in the 1970s [10, 11].
independent research fields with little overlap.
Over the past 30 years, high-resolution NMR at high
Having this in mind, it is interesting to recall that magnetic fields has not only become a routine method for
P URCELL and BLOCH (Nobel prize for Physics in 1952) molecular structure determination in Chemistry and
performed their first experiments independently of each other Pharmaceutics, but has also gained outstanding importance
on a solid and a liquid sample, respectively, namely paraffin for the characterisation of complicated biological systems, in
and water using proton (1H) resonances [1, 2]. In a sense, particular proteins. Rumour has it that BLOCH liked to say
NMR was discovered in parallel on solid and liquid it was him who he did the first biological NMR experiment
samples, and it was therefore clear from the very beginning in 1946 when he put his finger into the probe coil of his
that NMR would be applicable to both, liquid and solid nuclear induction apparatus and received a strong 1H NMR
phases of condensed matter. Despite this common start, signal from it. However, the first serious high-resolution
liquid-state and solid-state branches bifurcated in the NMR studies of living systems began with the phosphorus
following, which was somewhat initiated by the discovery spectra of intact red blood cells by M OON and R ICHARDS in
of chemical shifts and spin multiplets between 1949 and 1973 [12]. Subsequently, the new field of “biological NMR”
1951 by PROCTOR and WU [3], and PACKARD and was particularly promoted by the work of WüTHRICH on
ARNOLD [4-6]. When molecules can tumble rapidly and protein structure determination [13], which has become an
isotropically in a dilute solution, the interactions that important subject of world-wide research efforts in numerous
determine NMR spectra are reduced to their isotropic parts, high-field NMR centres. The versatility and significance of
i.e., isotropic chemical shifts and so-called J-couplings. high-resolution NMR is also reflected by two Nobel prizes
in Chemistry awarded to ERNST [14] and WüTHRICH [15].
*Address correspondence to this author at the Max-Planck-Institut für in 1991 and 2002, respectively.
Polymerforschung, Postfach 3148, D-55021 Mainz, Germany; Tel: +49
6131 379-240; Fax: +49 6131 379-100; E-mail: schnelli@mpip- While liquid-state NMR has developed into a routine
mainz.mpg.de characterisation method with numerous applications in

1573-4110/05 $50.00+.00 © 2005 Bentham Science Publishers Ltd.


4 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

diverse fields of science and medicine, one might be tempted In view of this background, the work described in this
to say that solid-state NMR spectroscopy has remained a review aims at a thorough methodological combination of
delicate, though powerful, tool for specialists. Technically, liquid-state and solid-state NMR by merging concepts from
this is predominantly due to the strong anisotropic either side. Based on this strategy, new MAS NMR
interactions present in solid materials, which make higher methods and experiments have been developed for organic
demands on NMR techniques and give rise to broad and supramolecular and biomolecular systems. These classes of
complex lines in solid-state NMR spectra. A groundbreaking materials not only gain more and more importance in current
idea how to deal with these anisotropic interactions came research, but also clearly show that future NMR methods
from ANDREW [16] and LOWE [17] who independently will need to be applicable to samples which are neither rigid
invented magic-angle spinning (MAS) already in the late solids nor isotropic liquids. Thus, it is about time that
1950s. Making use of the orientation dependence of NMR spectroscopy overcomes the “traditional” separation
anisotropic interactions relative to the magnetic field (B0), between liquid state and solid state and brings its common
the sample is spun rapidly about an axis which is inclined at roots back to its mind.
the so-called magic angle relative to B0. In this way,
anisotropic parts of interactions are averaged, provided that
the spinning frequency exceeds the strength of the 2. INTRODUCING LIQUID-STATE NMR CONCEPTS
interaction. Consequently, the line narrowing performance is INTO SOLID-STATE MAS NMR
in many cases limited by the technically achievable MAS
frequency, which is why it was not until the 1990s that In NMR, interactions become orientation dependent due
sufficient spinning frequencies [18, 19] paved the way for to the presence of a strong external magnetic field, B0. This
widely spread applications of solid-state MAS NMR field gives rise to the famous ZEEMAN interaction, which
experiments including protons [20, 21]. usually exceeds all internal interactions by several orders of
magnitude. Consequently, the latter are treated as a first-
In 1998, equipment supporting MAS frequencies in order perturbation of the ZEEMAN interaction in the so-called
excess of 30 kHz became commercially available. In “rotating frame” [29], and the B0 field effectively represents
combination with the technical progress of radio-frequency the quantisation axis of the spin system. Based on this, two
circuits and pulse generators, solid-state NMR equipment classes of interactions can be distinguished, namely
has meanwhile reached a status which allows for an efficient anisotropic and isotropic interactions, which exhibit and do
handling and averaging of anisotropic interactions (apart not exhibit, respectively, an orientation dependence relative
from strong quadrupolar couplings, though). It should to B0.
therefore be possible to design solid-state NMR experiments
by incorporating techniques and concepts from liquid-state The fundamental difference between solid- and liquid-
NMR, which were so far supposed to be dedicated to the state NMR can also be traced back to this distinction, as it
liquid state only. Thus, it is time for solid-state NMR arises from the absence and presence, respectively, of rapid
spectroscopists to remind themselves of the wealth of liquid- isotropic motions of the molecules under investigation. In
state NMR techniques. this context, “rapid” refers to the timescale of the
interactions which are averaged and, thus, reduced to their
Interestingly, the same also applies in reverse. In the last isotropic properties by the motion of the molecules.
years, high-resolution NMR has taken substantial advantage Typically, NMR interactions give rise to energy level
of slightly aligning the molecules under inspection, for differences and transition frequencies of the order of ∆E/h =
example by dissolving them in a dilute liquid-crystalline 100 ... 2·104Hz [29], when large quadrupole couplings are
medium. In this way, small residual anisotropic interactions ignored. Correspondingly, molecular motions need to occur
are re-introduced and become available for structural isotropically with correlation times of τ C < 10µs, in order to
investigations of (large) molecules, in particular proteins. ensure complete averaging of anisotropic properties. This is
This strategy, pioneered by B AX [22, 23] and PRESTEGARD usually easily fulfilled in diluted solutions of relatively
[24, 25] has attracted considerable interest in the biological small molecules. Therefore, this class of samples constitutes
NMR community and undoubtedly bears further potential the classical field of application of liquid-state (or high-
for high-resolution NMR applications in general. Another resolution) NMR spectroscopy.
field of current interest is focused on materials in which the
molecules of interest can move rapidly, but not fully In solids, obviously, molecules do not undergo such
isotropically, because they are attached to an immobile rapid and isotropic motions, and NMR has to deal with
species. In addition to the motional restriction, such pronounced effects arising from anisotropic interactions
inhomogeneous mixtures of mobile and immobile [30, 31], such as anisotropies of the chemical shielding
components usually exhibit large magnetic susceptibility (CSA) as well as dipole-dipole and quadrupole couplings,
gradients which broaden the resonance lines and destroy the just to mention the most prominent ones. These effects
spectral resolution. In order to remove these unwanted dominate typical solid-state NMR spectra and make them
susceptibility gradients, together with other residual appear fundamentally different from high-resolution spectra
anisotropies, and to restore spectral resolution, MAS has obtained from solutions. Thus, the transfer of concepts from
recently been successfully introduced into high-resolution liquid-state NMR into its solid-state counterpart relies on
NMR spectroscopy, and the approach has been termed techniques that allow for an efficient averaging or, at least,
“HRMAS” spectroscopy [26-28]. Considering HRMAS and handling of anisotropic properties. In the past five years,
the strategy of partial alignment outlined above, it is significant advances into this direction have become possible
palpable that liquid-state NMR is about to take up concepts by the technical progress in the field of fast magic-angle
from solid-state NMR. spinning (MAS), which will be discussed in the first part of
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 5

this section. Consecutively, ideas will be introduced which are of practical importance in NMR research, because they
enable the selective restoration, i.e. “recoupling”, of certain can be used to selectively replace protons at sites of interest.
anisotropic properties. In this way, coherent spin states can In the secular approximation, the Hamiltonians of the
be generated, and numerous experimental schemes can be relevant anisotropic NMR interactions can be expressed in
designed, in which the strength of the anisotropic interaction terms of operators, which are components of rank two and
of interest becomes measurable through the coherent order zero of irreducible spherical tensor operators [34] where
evolution of spin states. Measurement strategies of this kind λ denotes the type of interaction, while i refers to the spin i
form the basis of a whole wealth of experiments in high- in case more than one is involved. For CSAs, hetero- (DIS )
resolution NMR spectroscopy, and the scope of this work is and homonuclear (Dij) dipole-dipole and quadrupole
to introduce these concepts into solid-state NMR couplings (Q) one finds[30, 31] for the space parts (given in
experiments. the principal axes system of the interaction, PAS), and for
the spin parts. The space parts consist of the respective
2.1. The Quest for Spectral Resolution coupling constants plus scaling factors, which deviate from
the usual conventions [30] because normalisation factors of
The impact of high-resolution NMR spectroscopy in the spin parts have been incorporated for reasons of clarity.
modern science can largely be attributed to its ability to The spin parts differ considerably for the interactions, but
sensitively distinguish different species of a nucleus, such as analogies are obvious between CSA and heteronuclear
1H, 13C, 15N etc., due to their chemical shifts. Being an dipole-dipole couplings as well as between homonuclear
isotropic property, these shifts become very well resolved dipole-dipole and quadrupole couplings.
when the molecules tumble rapidly and isotropically in
Hλ = Σ A λ2,0, i T 2,0
λ, i
solution. In solids, resonance lines are severely broadened by , (1)
i
anisotropic interactions, which pose the first and most
important obstacle to be overcome before considering
= δ , A DIS ,P AS = 2DIS , A D I J , PA S = 3D, ,ij
CSA .PA S
analogies between liquid- and solid-state NMR. A 2,0 2,0 2,0
Isotropic and Anisotropic Interactions Q ,P A S
= 3
δQ (2a)
and A 2,0 2
The concepts of liquid-state NMR spectroscopy are
largely built on the two isotropic interactions, namely the (i) (j)
= 2IZ S Z , T ij = 3IZ I Z I (i) I (j) ,
CSA D D
isotropic chemical shifts, which provides spectral resolution, T 2,0 = IZ , T IS
2,0 2,0
and the indirect spin-spin (or J-) couplings, which reflect the Q
network of chemical bonds [29]. In principle, these two and T 2,0= 3I Z I Z I I (2b)
interactions do not hamper spectral resolution, as they lead
to sharp resonance lines, although multiplet splittings Averaging Techniques
arising from J-couplings might confuse and complicate the To remove an anisotropic interaction, i.e., to average the
interpretation of the spectra. To accomplish spectral respective Hamiltonian (see Eq. 1), it is sufficient to cancel
resolution in solid-state NMR spectra, line broadening either the spin part or the space part. Formally, all averaging
effects of anisotropic interactions need to be effectively procedures involve a transformation of reference frames into
removed, while retaining the isotropic chemical-shift the laboratory system (LAB) with B0 || z. In other words, the
information. orientation dependences of the anisotropic interactions result
Basically, this work is restricted to spin-1/2 nuclei, in from the transformation of the reference frames into the LAB
particular to 1H, 13C and 15N due to their fundamental role frame. Generally, concerning the space part, this
in biomolecules. From a methodological point of view, this transformation occurs from the PAS to the LAB frame and
reduces the relevant anisotropic interactions to homo- and can be expressed as a rotation using the reduced
heteronuclear dipole-dipole couplings and chemical shifts, elements d (2)
m,0 (β) of WIGNER rotation matrices [30,31],
while quadrupole couplings can be ignored. With respect to where α and β are the polar and azimuthal angles specifying
the size of the interactions, this means a considerable the orientation of the interaction tensor. Eq. 3 can be applied
simplification, because quadrupole couplings can be very analogously to the spin operators. In spin space, rotations
strong and reach in some cases the same order as the are effectively accomplished by pulsed radio-frequency (RF)
ZEEMAN interaction. Then, they can no longer be considered fields that are applied at the resonance (i.e. LARMOR)
within the so-called secular approximation as a “weak” first- frequency of the respective nucleus. Then, the angles α and
order perturbation of the dominating ZEEMAN interaction, β represent the phase and the flip angle, respectively, of the
and it becomes technically very demanding (if not RF pulses [30].
impossible) to reduce them efficiently [32, 33]. Therefore, it λ, P AS
A λ,2,0LA B = Σ A 2,m
-ima (2)
is rather difficult to propose general concepts of merging e d m.0 (β) (3)
m
liquid- and solid-state NMR including nuclei with strong
quadrupole couplings. An important exception is the All averaging schemes, acting either in real space or in
deuteron, 2H, because its typical quadrupole couplings spin space, are formally based on Eq. 3. To achieve
hardly exceed 200 kHz and are thus only by a factor of < 10 averaging, α is effectively made time dependent by
stronger than the strongest dipole-dipole couplings. introducing ω t. This results in harmonic oscillations,
Therefore, they can be treated fully analogously to dipole- exp(+-imω t), which average to zero over full oscillation
dipole and chemical-shift interactions. Moreover, deuterons periods. In addition, β is chosen such that the static non-
oscillating component (m = 0) disappears as well, which is
6 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

the case for the so-called “magic angle” βm = 54.7°. as well as by numerically optimised pulse schemes [39].
Therefore, the per-formance of such averaging techniques Such continuous RF schemes are advantageous insofar as
crucially depends on the precision of the magic-angle high averaging frequencies of the order of 100 … 200 kHz
adjustment and on the frequency ω . With respect to the can be realised experimentally. However, the observed
latter, it is important to recall the distinction between isotropic chemical shifts are effectively scaled by factors
homogeneous and inhomogeneous line broadening < 1 3 [30], which significantly reduces the separation of
phenomena, as introduced by MARICQ and WAUGH [35]. the resonance lines in the spectra. Moreover, in order to
Theoretically, inhomoge-neous interactions, such as CSAs, detect signal or to apply other RF pulse sequences at the
heteronuclear dipole-dipole or quadrupole couplings, are same time, the RF averaging schemes need to be interrupted,
fully averaged over one oscillation period irrespective of the which usually reduces their efficiency and severely
frequency, while the homogeneous character of homonuclear complicates the RF pulse programming. MAS, in contrast,
dipole-dipole couplings requires the period of the averaging only allows for averaging frequencies of the order of 30 kHz
cycle to be short enough to prevent the formation of a (at the present stage), but it leaves the RF side of a NMR
dipolar-coupled multi-spin network. In practice, pure spectrometer completely free and available for pulse
inhomogeneous line broadening is rarely found, because sequences of NMR experiments. In a sense, MAS mimics
homogeneous effects are encountered as soon as homonuclear the rapid isotropic motions the molecules undergo in dilute
dipole-dipole couplings are present in the spin system. This solutions, and paves the way for the introduction of liquid-
is usually the case, as most systems of interest contain a state techniques into solid-state NMR. Due to this
significant amount of protons with multiple strong dipole- conceptual analogy, the work described here concentrates on
dipole couplings among them. Therefore, the averaging MAS and its ability to average anisotropic interactions.
frequency ω , in general, needs to be high enough to Resolution Enhancement by Magic-angle Spinning
efficiently suppress proton-proton dipolar interactions, i.e.,
ω /2π ≥ 25 kHz as a rule of thumb. For most (bio)organic spin-1/2 systems, the spectral
resolution achievable in solid-state NMR is determined by
As mentioned above, the averaging procedures can the ability to suppress dipole-dipole couplings to and among
formally be applied in real space as well as in spin space. In protons, i.e., to “decouple” protons. Therefore, simple 1H
real space, magic-angle spinning (MAS) is the most NMR spectra recorded at different MAS frequencies can
commonly used technique which involves a rapid mechanic serve to illustrate the decoupling performance of MAS,
rotation of the sample about an axis that is inclined at the which is shown in (Fig. 1) using a crystalline benzoxazine
magic angle relative to the B0 field [16, 17]. In spin space, dimer as an example [20].
the same type of rotation is accomplished by the application
of suitable RF fields. Presently, maximum performance is It is obvious from (Fig. 1) that, for 1H spectra of rigid
provided by LEE-GOLDBURG pulse schemes [36], their solids, MAS needs to be applied at frequencies ≥ 25 kHz. At
frequency-switched [37] and phase modulated [38] variants higher spinning frequencies, the resolution still increases

Fig. (1). Spectral resolution achievable at various MAS frequencies in the 1 H solid-state MAS spectrum of a crystalline benzoxazine
dimer (the chemical structure of which is depicted in the inset).
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 7

gradually, but the maximum resolution is not only πJ IZ SZ


IX IX cos(πJt) + 1Y SZ sin (πJt), (4)
determined by the technically available MAS frequencies,
but also by the material itself. When the crystallinity, i.e., In view of this successful concept, the question arises
the structural order, of a material is not perfect, the local whether and to what extent the idea of generating and
distributions of electrons around the nuclei are affected by transferring coherent two- or multi-spin states would also be
local disorder or variations of the packing structure. These applicable under solid-state conditions. Obviously, in
phenomena give rise to dispersions of the chemical shifts solids, anisotropic interactions and the associated line-
and, thus, to an inherent line width which cannot possibly broadening and relaxation effects represent the main obstacle.
be reduced by averaging techniques. Therefore, in amorphous However, if averaging techniques can reduce these
samples, maximum 1H resolution is already achieved at anisotropies efficiently, and unravel the isotropic
MAS frequencies of about 30 … 35 kHz, and faster spinning information, similar experiments become possible in solid-
would not further increase the spectral resolution. state NMR as well. For this purpose, the recovery of the
Despite this limitation, the quest for higher MAS isotropic chemical shifts is no doubt essential, but with
frequencies [40] is still perfectly legitimate, because the regard to the J-couplings, there is an alternative way to
decoupling of the dipolar-coupled proton network is not couple nuclei in solid samples, namely via dipole-dipole
only required to enhance resolution in 1H spectra, but also to couplings. Having said this, three major differences between
reduce transverse relaxation phenomena of other nuclei X, dipole-dipole and J-couplings are obvious:
such as 13C or 15N. Usually, such heteronuclear decoupling (i) J-couplings are electron-mediated indirect spin-spin
between X and 1H is accomplished by powerful RF pulse couplings which are, as a consequence of this,
schemes [41-44] which are applied on the 1H channel while restricted to nuclei linked by chemical bonds. Dipole-
observing the X signal. This simple and efficient approach, dipole couplings, in contrast, act directly through
however, provides full decoupling between 1H and X nuclei, space and only depend on the distance, r, between the
which is no longer feasible when correlated spin states of the nuclei according to 1/r3. Therefore, they are not
form X-H are to be observed. Then, a decoupling scheme is directly related to the network of chemical bonds,
required which does not destroy the correlated X-H spin state which on the one hand hampers the investigation of
(as brute-force RF decoupling would do), but efficiently the chemical structure of molecules, but on the other
prevents transverse relaxation. This issue gains importance hand enables the detection of intermolecular contacts.
for so-called inverse detection schemes, which are discussed
in the following section. (ii) For the nuclei discussed here, J-couplings are
basically fully isotropic interactions, while dipole-
dipole couplings are fully anisotropic and do not
2.2. The Creation of Coherent Spin States possess an isotropic component at all.
Aiming at the removal of anisotropic interactions, the (iii) The two types of couplings also differ considerably
previous considerations were mainly concerned with with respect to coupling sizes: Heteronuclear 15N-1H
restoring the information on isotropic chemical shifts. There and 13C-1H J-couplings are of the order of
is, however, another isotropic interaction, namely the 80 … 150 Hz, and typical homonuclear 1H-1H J-
indirect spin-spin (or J-) coupling, which is of outstanding couplings do not exceed 15 Hz. The dipolar
importance in high-resolution NMR spectroscopy, because it counterparts are of the order of 10 … 20 kHz, and
enables the correlation of spin states and the creation of thus larger by factors of about 102 … 10 3. This
multi-spin coherences [10, 29]. In this way, J-couplings comparison, however, only holds in the absence of
form the basis for virtually all multidimensional NMR (molecular) motions, because dipole-dipole couplings
experiments in the liquid state. are averaged and reduced by molecular dynamics
and/or mechanical sample rotation (MAS) due to their
In the presence of a J-coupling, represented by a anisotropic nature.
Hamiltonian of the form HJ = πJ . Iz S z , a two-spin system
(I,S) oscillates between a correlated (IYS Z) and an At first sight, the significant differences between dipole-
uncorrelated, transverse spin state (IX), which can be dipole and J-couplings seem to suggest that a transfer of
expressed as [10] where J is the J-coupling constant. When experimental schemes from liquid- to solid-state NMR relies
observing the I-spin signal, the cosine-modulation gives rise on techniques which provide access to J-couplings in solids.
to the well-known multiplet splittings in the spectra [10,29]. As J-couplings are very weak compared to typical
In view of multidimensional NMR, however, the correlated anisotropies encountered in solids, powerful averaging
spin state is the more interesting one, as it provides a means techniques are required, which make quite high demands on
to coherently transfer states from one spin to another. In the the technical equipment in terms of available RF power and
case of J-couplings, such transfer processes occur between RF stability. Such approaches exist, pioneered by LESAGE
chemically bonded spins, which means that spin states are and EMSLEY [45-49], but at the present stage, only RF-
propagated through the molecule via the network of chemical based decoupling schemes in combination with MAS are
bonds. Ideally, due to the coherent nature of the processes capable of providing sufficient averaging of anisotropic
and the (formal) absence of relaxation, no information is lost interactions. At the current limit of spinning frequencies,
in the course of the spin system’s evolution. This direct link MAS alone does not suffice to unravel J-couplings in solids.
between spectroscopic properties and molecular structure Nevertheless, the solid-state NMR methods presented
represents one of the fundamental reasons for the success and herein do not consider J-couplings, but are solely based on
versatility of high-resolution NMR spectroscopy. dipole-dipole couplings because of two reasons: Firstly,
8 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

dipole-dipole couplings are much stronger, and it is therefore λ.PAS


A 2,0
much easier to use them in the experiments rather than Φλ (t0;t1) = 1 sin2 β(sin (2ω t +2γ) -sin(2ω t + 2γ)
ωR 2
R1 R0
battling against them with sophisticated pulse sequences.
Secondly, dipole-dipole couplings are efficiently averaged by 2 sin 2β (sin (ωRt1 + γ) - sin (ωRt0 + γ) (6)
fast MAS, but can conveniently be re-introduced by so-
called recoupling pulse sequences, which are discussed λ
HMAS
below. In this way, the experiments gain an important ρ0 ρ0 cosΦ λ + W sin Φ λ (7)
degree of flexibility, because dipole-dipole couplings can be
switched on and off during the experiments basically at will.
During recoupling periods, coherent dipolar two- and multi- Recoupling Schemes
spin states can be generated, and concepts of high-resolution The MAS NMR experiments considered in this part of
NMR experiments can be adapted to solid-state MAS the work employ sequences of strong RF pulses separated by
conditions, including coherence transfers. Even more, the delays, during which the spin system evolves under the
peculiarities of MAS and recoupling procedures allow for an internal interactions. These pulse sequences are often referred
extension of such concepts towards sensitive measurements to as “laboratory frame” sequences, because the quantisation
of couplings. As a starting point, however, the effect of axis is given by the static magnetic field B0 along the
MAS on anisotropic interactions needs to be considered first laboratory z-axis. “Rotating frame” pulse sequences, in
in more detail. contrast, are characterised by a continuous application of RF
Rotor-Modulated Interactions fields, so that the spin system evolves no longer freely but
in the presence of transverse RF fields (see for example the
Under MAS, the effective Hamiltonian of an anisotropic helical pulse sequences discussed in Chapter 3).
interaction represented by a second-rank tensor can be
approximated using the zeroth-order average Hamiltonian The principle of laboratory-frame recoupling pulse
[30, 50] sequences is well established [51, 52] and can easily be
understood by considering the rotor modulations which are
λ λ
t1 λ LA B introduced by the sample rotation (see Eq. 6). The spatial
A ,
1
t0 - t1 t∫ 2,0
HM AS (t0 ; t1) = T 2,0 (ωRt)dt
parts of the interaction Hamiltonians are modulated by the
MAS frequency ω R and twice the MAS frequency 2ω R. For
0
λ
= T2,0 t - t Φ λ (t0 ; t1)
1
(5) illustration, the respective oscillations are depicted in (Fig.
0 1
λ λ
2b). By averaging – in other words: integrating – over a full
Α L, M and Τ L,M represent the spatial part and the spin rotor period, the modulated interactions vanish because
part (see Eq. 2a and 2b) of the interaction λ, and are given as positive and negative parts of the oscillations cancel each
components of irreducible spherical tensor operators of order other. This cancellation effect can also be seen from Eq. 4,
M and rank L [30,34]. The first term with L = M = 0 is when t1 – t0 = n⋅τ R and n is an integer number. An obvious
neglected, as it represents the isotropic part of the way to avoid this cancellation is to invert the sign of the
interaction, which is zero for dipole-dipole and quadrupole spin part by a pulse sequence simultaneously to the sign
interactions: Α0.0 = ΑQ λ
0.0 = 0 The integrated phase Φ (t 0;t 1)
D
changes imposed on the spatial part by MAS. Effectively,
contains the spatial part of the interaction, which is recoupling is accomplished by inducing a “counter-rotation”
modulated by the sample rotation [30] where Α λ.P 2.0
AS
is the in spin space relative to MAS rotation in real space. As there
relevant tensor component in the PAS of the interaction λ are two modulation frequencies, ω R and 2ω R, the sign
(as given explicitly in Eq. 2a), and ω R denotes the MAS change in the spin part can be performed in intervals of
frequency. β and γ are the azimuthal and polar angles that either τ R/2 or τ R/4 [93].
describe the orientation of the interaction tensor in the rotor
frame (see Fig. 2a). γ can also be viewed as the initial rotor The type of RF pulses required for this sign inversion
λ
phase, as the time-dependent rotor phase, ω Rt, is always depends on the nature of the spin part T2,0 of the respective
added to this angle. In Eq. 6, it is assumed that the interaction λ (see Eq. 2b). For interactions depending
interaction is symmetric, which greatly simplifies the linearly on a spin (i.e., CSA and heteronuclear dipole-dipole
description in the case of dipole-dipole couplings and many couplings), the sign inversion is straightforwardly achieved
2H quadrupole couplings. For CSAs, asymmetries need to by π pulses. The approach of utilizing π-pulse trains with
be taken into account, which requires the tensor components pulse spacings of τ R/2 was first realised in the REDOR
ΑCSA
2,+2
. PAS
to be included. Using the above expressions for (rotational echo, double resonance) experiment, which was
an averaged Hamiltonian under MAS, an initial state of initially introduced by SCHAEFER and GULLION [53-56] in
transverse magnetisation, ρ0 = Ix (ρ 0 = I x(i ) + Ix(j) ) for a spin order to recouple the heteronuclear dipole-dipole coupling
pair ij with a homonuclear dipole-dipole coupling), evolves between two kinds of low-γ nuclei. Such REDOR-type π-
under an anisotropic interaction according to where W pulse trains are also commonly used for CSA recoupling
denotes the spin state into which the system evolves under [57, 58] or for recoupling 1H-X heteronuclear dipole-dipole
the interaction λ, i.e. W = IY, W = 2IYS Z, couplings, 51, 52, 55, 56] where X denotes a low-γ spin-1/2
W = 2(IY(i) I(j) I(i) I(j) ) and W = 2(IYIZ + IZIY) for nucleus. Interactions with a bilinear spin dependence (i.e.,
Z Z Y
CSA, hetero- and homonuclear dipole-dipole and quadrupole homonuclear dipole-dipole and quadrupole couplings) can be
coupling, respectively. Eq. 7 represents a general form of recoupled in an analogous fashion by the so-called Back-to-
Eq. 4, which has been adapted to rotor-modulated Back (BABA) pulse sequence [21, 59]. In this sequence,
anisotropic interactions, as encountered in solids under MAS each π-pulse of REDOR-type recoupling is replaced by a
conditions. pair of π/2-pulses with orthogonal phases. Effectively, these
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 9

Fig. (2). (a) Geometric relations under MAS conditions, including the definition of the angles β and γ. They relate the main axis of a
tensor representing a symmetric anisotropic interaction to the rotor. (b) Modulations of an anisotropic interaction under MAS (top)
and pulse sequences for recoupling interactions which depend on the nuclear spin in a bilinear (middle) or linear fashion (bottom).
The pairs of 90° RF pulses (middle) recouple homonuclear dipole-dipole couplings and (weak) quadrupole couplings, while the 180°
pulses (bottom) recouple heteronuclear dipole-dipole couplings or CSAs.

λ
pairs of π/2-pulses invert the sign of the spin part T 2.0 of the same signal amplitude as ω R-recoupling. Consequently,
the homonuclear dipole-dipole and quadrupole coupling. ω R-recoupling is in general favourable with respect to the
recoupling efficiency, and 2ω R-recoupling is disadvanta-
In Fig. 2b, the principles of BABA and REDOR
geous unless sample orientation becomes an issue, which
recoupling are illustrated [93]. Clearly, pulse trains with
will be discussed below.
pulse spacings of τ R/2 recouple only the ω R-modulated
terms, while pulse trains with pulse spacings of τ R/4 Two-Spin Correlations and Double-Quantum Coherences
recouple only the 2ω R-modulated terms. Henceforth, these The recoupling procedures outlined above allow for the
two types of recoupling will be referred to as “ω R- generation of states W according to Eq. 7, where the
recoupling” and “2ω R-recoupling”. [93]. These experiments integrated phases Φ λ are to be replaced by the recoupled
give rise to the following integrated phases for axially λ
phases NR Φ nω R (t) (with n =1 or 2) given in Eqs. 8a and
symmetric interactions [93], 8b. N R denotes the number of rotor periods, during which
2 Φ λ (t;t + τR /2) = Φ ωλ (t) recoupling is applied (i.e., the recoupling time), while t
R defines the starting time of the recoupling period in the
Aλ,2,0PA S experiment, which reflects the so-called rotor encoding of the
= ω 2 sin 2β sin (ωRt + γ) (8a) recoupled interaction [21, 60] and will become crucial for the
R
generation of sideband patterns (see section 2.3). Before
λ
4 Φ λ (t;t + τ R /4) = Φ2ω (t) = turning to these, the state W deserves some discussion.
R
λ, PA S As mentioned above, W can take different forms.
A 2,0 Evolving out of an initial state ρ0 = IX, W remains a single
= ω sin β sin (2ωR t + 2γ)
2 (8b)
R spin state, when the (recoupled) interaction is not a spin-spin
coupling, but instead represents the anisotropy of the
The factors 2 and 4 have been introduced to make the
chemical shielding (CSA) or the quadrupole coupling
total timing of both expressions equal (i.e. τ R) and to allow
between the nuclear quadrupole moment and the electric field
for a comparison of their absolute scaling. Then, it becomes
gradient at the nucleus. This can directly be seen from the
obvious that the two expressions differ not only in their
Hamiltonians, which contain operators of a single spin only.
orientation dependencies (sin 2β and sin 2 β), but also in their
In spin-1/2 cases (CSA), the spin simply stays in a
amplitudes. Under ω R-recoupling, the effective Hamiltonian
transverse state, the phase of which is modulated by the
is larger by a factor of 2 as compared to 2ω R- recoupled CSA. In the terminology of coherences, such
recoupling, which means that, theoretically, 2ω R-recoupling
transverse states correspond to single-quantum coherences
needs to be applied longer by a factor of, 2 to achieve
(SQCs), because the spin state can be understood as a
10 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

coherent superposition of the states with magnetic spin Procedures of this kind are well established as building
quantum numbers mI = +1/2 and mI = –1/2. This involves a blocks of liquid-state NMR experiments [10], and they can
transition of |∆mI | = 1, which is a SQ transition. also be realised in solid-state experiments by combining the
averaging performance of fast MAS with suitable recoupling
In the quadrupolar case (i.e., nuclei with spin ≥ 1),in
pulse sequences. As the transfer occurs coherently, the
contrast, the spins can evolve into coherences of higher order
amplitude of the final state (see Eq. 9) contains the full
under the quadrupole coupling. As this work is only
information on the heteronuclear dipole-dipole coupling.
concerned with 2H as a quadrupolar nucleus, the discussion
Thus, coherent transfers not only occur in a well defined
can be restricted to the spin-1 case with relatively weak
manner, they also enable the measurement of the involved
quadrupole couplings. The only higher-order coherence
coupling, as will be discussed below. In these respects, they
available to deuterons is the double-quantum coherence
are superior to incoherent transfer processes, such as the
(DQC) which involves a transition of |∆mI | = 2. At first
well-established technique of “cross polarisation” in solid-
sight, it might appear a bit surprising that in NMR theory
state NMR [30].
such DQCs are commonly considered although quantum
mechanics says that the DQ transition is forbidden. In
NMR, however, this rule does not apply because the spin 2.3. The Measurement of Couplings
states under consideration are non-equilibrium states that
have been created by a strong perturbation of the spin system After describing how and which coherent spin states can
(i.e., the application of a high power RF pulse) [29]. be generated in solid-state MAS NMR experiments by
Consequently, in the case of deuterons W represents a DQC recoupling anisotropic interactions, it is now time to turn to
that is generated through the recoupled quadrupole coupling. the actual purposes of all these efforts, namely to
experiments which allow anisotropic interactions to be
When spin-spin couplings, such as J- or recoupled
measured in a selective and sensitive fashion. These
dipole-dipole couplings, act on the spin system, W
measurements aim to provide detailed insight into the
represents two-spin states, which again directly reflects the
structure of molecules, their packing and their dynamics. In
properties of the Hamiltonians. Compared to the single-spin
this respect, high-resolution NMR spectroscopy has already
scenarios described above, the two-spin interactions provide
proven its enormous versatility for characterising solubilised
access to two- and multi-spin correlations and, subsequently,
systems, and the goal of this work is the design of
to coherence transfer processes. A homonuclear two spin-1/2
experiments which enable solid-state NMR to unearth as
system with a dipole-dipole coupling bears analogy to a
much information as possible about solid materials, which
spin-1 quadrupolar nucleus insofar as the coupled two-spin
is so far buried in the spectra by solid-state effects.
system can form a DQC. In contrast to the 2H case,
however, the dipolar DQC is, in general, not the highest All solid-state NMR experiments discussed in this
possible coherence order, because further spins can couple to review, in principle, follow the same scheme, which is
the DQC and, in this way, produce multiple-quantum depicted in (Fig. 3). While fast MAS is continuously
coherences of various orders [61, 62]. When the number of applied, anisotropic interactions, primarily dipole-dipole
spins and couplings increases, the coherent nature of the spin couplings, are recoupled by appropriate pulse sequences
state becomes more and more concealed and is finally lost. during two periods. These are separated by an indirect
In other words, such multi-spin effects bridge the gap spectral dimension t1, which is inserted to shift the phases
between coherent spin states and incoherent relaxation of the two recoupling blocks relative to each other and/or to
phenomena. record a spectrum with chemical-shift resolution. In some
cases, the latter can alternatively be achieved by placing a
In a heteronuclear two spin-1/2 system, the situation is
spectral dimension t1 before the first recoupling block.
simplified by the large difference in LARMOR frequencies
Optionally, the experiment starts with a preparation period
between the two spins involved. As a result, the spin part of
during which the initial state of the spin system required for
the Hamiltonian is truncated (see Eq. 2b) [30], and the spins
the experiment is prepared; for example, transverse 13C
can separately be manipulated by RF pulses. Therefore, in
magnetisation can initially be prepared by a conventional
the experiments, an initial state ρ0 = IX evolves first into a
cross polarisation of 1H magnetisation. At the end of all
two-spin correlation, or “antiphase coherence”, of the form
pulse sequences, the signal is detected during the direct
W = 2IYS Z, which does not evolve into a DQC unless an
spectral dimension t2.
additional RF pulse is applied on the S-spins. More
generally speaking, in heteronuclear experiments, RF pulses Rotor Encoding and Sideband Patterns [93]
can be used to switch between transverse (IX,Y, SX,Y) and An efficient and sensitive way to measure anisotropic
longitudinal spin states (IZ, SZ) independently on the two interactions is the generation of spinning sideband patterns
spins involved. In this way, spin states, such as transverse via rotor encoding (RE) of the recoupled interactions [21,
magnetisation, can straightforwardly be transferred from the 60]. Such rotor encoding is accomplished by combining two
I- to the S-spin through a transfer sequence of the form recoupling blocks with different initial rotor phases,
DIS meaning that the second block is shifted with respect to the
H MAS (π2) I (π2) S first by a time t1 which is incremented in steps ∆t1 < τ R
IX 2IY SΖ sin Φ λ 2IΖ SY sin Φ λ (see Fig. 3). The amplitude of the recoupled signal is then
modulated (for ω R-recoupling) according to [93] depending
DIS
H MAS on the coherence pathway selected and observed in the
SX sin Φ λ sin Φ λ (9) recoupling experiment. For 2ω R-recoupling, ΦωλR would
have to be used instead of ΦωλR (see Eqs. 8a and 8b). The
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 11

Fig. (3). Schematic representation of the two-dimensional recoupling experiments described in this work.

time t1 represents the delay between the two recoupling obvious from Eqs. 6 and 8 that, for ω R- and 2ω R-
blocks, and N exc and N rec denote the numbers of rotor recoupling, full echoes occur in the t1 signal every full and
periods during which the first and second recoupling half rotor period, respectively. Therefore, after Fourier
sequence is applied (referring to “excitation” and transformation along t1, spinning sidebands are, in principle,
“reconversion”, respectively). Following Eq. 7, the found at all multiples of ω R in the ω R-recoupled spectra,
observation of the initial state ρ0 would give rise to a signal while 2ω R-recoupling leads to sidebands only at multiples
of the form Scos (Eq. 10a), while the evolution into a state of 2ω R. More specifically, a signal of the form Ssin,cos gives
W and back into observable magnetisation would yield a rise to sidebands at all multiples of ω R and 2ω R (for ω R-
modulation of the form Ssin (Eq. 10b, as also found for the and 2ω R-recoupling, respectively), while Ssin and Scos lead
transfer described by Eq. 9). In both cases, the spin states ρ0 to sidebands only at odd and even multiples, respectively, of
and W need to be selected between the two recoupling ω R and 2ω R [93]. An example for sidebands arising from
blocks, and other contributions need to be removed by an ω R- and 2ω R-recoupling of a 1H-13C dipole-dipole coupling
appropriate phase cycle of the RF pulse sequence. Without is given in Fig. 4. The signals and sideband patterns shown
such a selection procedure, a superposition of sin- and cos- there correspond to the Ssin-type (Eq. 10b).
type evolutions leads to a signal of the form Ssin,cos
according to Eq. 10c. It should be noted that this S cos (t1) ~ cos Ne xc Φωλ (0) cos Nre c Φωλ (t1 ) (10a)
R R
consideration is simplified insofar as any evolution of the
spin system (in a non-longitudinal state) occurring during
Ssin (t1 ) ~ sin N ex c ΦωλR (0) sin Nre c Φωλ (t1 ) , or (10b)
the short t1 intervals is neglected. “Pure” ω R- or 2ω R- R
recoupling without contributions from the other modulation
is only ensured when the spin state present during t1 is not
subject to evolution. Otherwise, the evolution during t1 S sin c os (t1 ) ~ sin Ne xc ΦωλR (0) + cos Ne xc ΦωλR (0)
gives rise to additional ω R- and 2ω R-modulations that enter
into the overall average Hamiltonian [93].
sin N rec Φωλ (t1) + cos N rec ΦωλR (t1) (10c)
While incrementing t1, the observed signal forms echoes R
when Φ λωR (tI ) = Φ λωR (0) or Φ λ2ωR (tI ) = Φ λ2ωR (0). It is

Fig. (4). Modulations of the t1 -signal (on the left), arising from rotor-encoding in a 1 H-13 C dipolar recoupling experiment performed
under ωR - and 2ωR -recoupling conditions. The corresponding sideband patterns are shown in the middle. The experimental data was
recorded on the CH resonance of powdered L-alanine at 25 kHz MAS. Under ωR - and 2ωR -recoupling conditions, the 1 H-13 C dipole-
dipole couplings were recoupled for 2τR and 4τR , respectively (see experimental scheme in Fig. 3). The dotted sideband patterns are
calculations using r CH = 113 pm. On the right, the dependence of the sideband patterns on the product of the dipole-dipole coupling
and the recoupling time is shown for a series of calculated patterns.
12 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

Directed Recoupling and Inverse Detection [63-67] the spin system can lead to a reversal of the role of I and S
spins. In the various types of heteronuclear correlation
In experimental pulse schemes, the recoupling sequences
experiments, different pathways of transverse coherences can
described above can be combined with spectral dimensions
be chosen [65]. The potential of this option lies in the
in various ways. The insertion of a t1 dimension between the
fundamental difference of the coupling topologies when one
two recoupling blocks has already been discussed with of the two spin species considered is isotopically dilute (or
respect to the rotor encoding and the sideband patterns spatially isolated). A transverse S coherence evolves as [65]
resulting from this. Of course, in many cases chemical shift while transverse I coherences evolve as [65] where Φ(i) =
resolution is also desired, which is provided by the spectrum ΦD IS.(i) or Φ D IS.(i) , depending on the type of recoupling.
ωR 2ωR
recorded in the direct dimension (t2, see Fig. 3) and can Comparing Eqs. 11 and 12, it becomes obvious that the
additionally be provided by an indirect dimension. In the structure of the modulation experienced by evolving I-spins
experiments discussed here, the correlation of two such is much simpler, because it consists of a sum of pair
spectral dimensions occurs via recoupled dipole-dipole couplings. When I-spins are transverse for the whole course
couplings, because they are capable of transferring spin states of the experiment, i.e., an II experiment is performed (see
from one spin to another. In homonuclear cases (e.g., 1H- Fig. 6c), the final signal is a superposition of pair
1H), two principal experimental schemes can be designed
signals [65].
(Figs. 5a and 5b), which differ in the spin state detected in
the indirect dimension: either a transverse single spin state,
i.e., a SQC, is detected before the dipolar transfer step (Fig.
5a), or a two-spin correlation, i.e., a DQC, is observed in
the middle of the (split) transfer sequence (Fig. 5b). The two
experiments therefore represent two-dimensional SQ-SQ or
DQ-SQ correlations [68], respectively.

Fig. (6). Coherence transfer pathways in hetero-nuclear dipolar


2D correlation experiments, referred to as (a) IS, (b) SS, (c) II and
(d) SI recoupling. (c) and (d) involve “inverse” detection.
(i)
Σ iΦ(i) IΖ SΖ
SX S γ Π i cos Φ (i)
(i)
Σi IΖ S γ sin Φ(i) Π j i cos Φ (i ) ... , (11)
=
Fig. (5). Homo- and heteronuclear dipolar 2D correlation while transverse Icoherences evolve as [65]
experiments resulting from different combinations of dipolar (i)
Σ iΦ (i) IΖ S Ζ
Σ i IX cos Φ
recoupling blocks and spectral dimensions. (i) (i) (j)
Σ i IX
For heteronuclear experiments (e.g., 1H-13C
or 1H-15N),
Σ i IY S Ζ sin Φ
(i) (i)
the presence of two distinguishable species of nuclei allows ... , (12)
for one more degree of freedom in the design of pulse
sequences, because during the recoupling periods each of the ~ Σ sin Ne xc Φ (i) (0)
SII sin Nre c Φ (i) (t1 ) , (13)
two spin species can separately be chosen to occupy a i
transverse state, as schematically depicted in Fig. 6. To
discuss this possibility and the consequences arising from it In the corresponding two-dimensional spectrum, the
in more detail, a simple spin system consisting of abundant detected I-spins can be resolved from each other by their
I spins and rare S spins will be considered in the following, chemical shift, while in the indirect dimension, in general,
as for example represented by a system with 1H and 13C in two options are possible: The two-spin correlation can be set
natural abundance under the application of a pulse sequence to a full longitudinal state of heteronuclear dipolar order
probing a heteronuclear two-spin state. It should be noted (HDOR, i.e., IZS Z) [65, 69, 70], which does not evolve at
that isotopic labelling schemes and a specific topology of all. As a consequence of this, pure rotor encoding leads to
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 13

sideband patterns, which may be a sum of two-spin patterns, This leads to sideband patterns which, in comparison to
because different I spins couple to different S-spins. the SS experiment, are even less influenced by the remote
However, if S-spin chemical-shift resolution is required, a couplings represented by the cosine terms. Only the first-
single-quantum antiphase state with a transverse S compo- order sidebands suffer from remote couplings to an
nent (i.e., IZS X,Y), can provide the necessary chemical-shift appreciable degree, and can be excluded from the analysis. It
separation also in the indirect dimension [65, 70]. should be noted that the one-dimensional version of REPT
is also known as TEDOR (transferred-echo, double-
Direct S detection (SS experiment, see Fig. 6b), in resonance, see also Fig. 5c) [77].
contrast, leads to signals of the type [65].
The SI recoupling scheme (inverse-detected REPT, Fig.
SSS ~ Σ sin Nexc Φ(i) (0) sin Nrec Φ(i) (t1) 5d) features essentially the same advantages and
i shortcomings as the IS experiment. The need for an initial
(i) (j) cross-polarisation step to transfer the initial magnetization to
Π cos Nexc Φ (0) cos Nrec Φ (t1)
j=i (14) the rare S nuclei and the subsequent TEDOR transfer back to
the abundant I-spins may at a first glance represent a serious
The additional cosine terms lead to problems in disadvantage as compared to the other three variants.
REDOR-based correlation experiments, as they introduce an However, it turns out that the SI experiment is the inverse
angular dependence into the experimental data and variant which is so far best applicable to samples naturally
complicate the interpretation [71, 72]. Corresponding abundant in 13C or 15N [64, 65].
interference effects are also known to arise from multiple J- Using II- and SI-type recoupling schemes, the concept of
couplings in liquid-state NMR [10]. There is even an inverse detection, which is well established and of great help
angular dependence which does, however, not result from an in high-resolution NMR [11, 108], can be adapted to solid-
orientation dependence of the J-couplings (as there is none), state conditions. The inverse, i.e., 1H, detection schemes not
but from the relative angles of chemical bonds [10]. only provide the flexibility discussed above for determining
An analysis of the spinning sidebands is also possible multiple couplings, but also and more importantly allow the
for an SS experiment when only the dominating couplings achievable signal-to-noise ratios to be significantly enhanced
are to be determined [70]. Then, the cosine terms in Eq. 14 by factors of 10 and more [63,64,66]. In this way, natural-
representing weak-coupling perturbations merely lead to a abundance 15N spectroscopy becomes feasible in solid-state
slight overestimation of the measured coupling values. A NMR [64, 65].
second coupling which reaches a similar magnitude as the Signal Selection and Suppression Using PFGs [78]
primary one renders the analysis of the pattern in terms of a
An important technical difficulty associated with inverse
single spin-pair critical. On including the second coupling,
(1H) detection schemes in 1H-X correlation spectroscopy is
however, the angle between the two involved coupling
the handling of excess 1H magnetisation. As the
vectors becomes accessible [65]. In fact, the sideband
heteronucleus X is usually much less abundant in the
patterns are surprisingly sensitive to the angle when
materials than 1H (for comparison: 1H 100%, 13C 1.1% and
“asymmetric” recoupling sequences, i.e., different excitation 15N 0.35% natural isotope abundance), the vast majority of
and reconversion times, are applied [65]. In the case of I2S 1H nuclei does not come into touch with the heteronuclei of
spin systems, for example, the large potential of directed
interest. Their intense signal contribution must therefore be
recoupling becomes appreciable. The individual coupling
removed from the detected signal or, preferably, be
constants can be determined precisely by performing an II
suppressed before the signal is detected. In conventional
experiment (which might not necessarily be an inverse-
phase cycling schemes of the RF pulses, unwanted signals
detected experiment, for example when the S spin is a
are cancelled by subtracting the signal of consecutive
proton), while the SS experiment can be used to determine
transients of the pulse sequence. This procedure, however,
the angle, using the known coupling constants as fixed
becomes the more disadvantageous the stronger the
parameters.
unwanted signal contributions are. This is because the
We are now left with the IS and SI recoupling schemes digital receiving units of the spectrometer are at some point
(shown in Figs. 6a and 6d), which involve a polarisation saturated with the unwanted signal, while the weak wanted
transfer (termed “recoupled polarisation transfer”, REPT) contributions fall below the digitisation threshold.
[69]. The IS experiment (Fig. 6a, with S being identified
In high-resolution NMR, this problem has been solved
with 13C in natural abundance) is conceptually simple
by introducing pulsed field gradients (PFGs), [79, 81]. For
because the pulse sequence does not contain a cross
a short time in the range of a few milliseconds (“pulse”),
polarization, and has already proven to be useful in a variety
additional magnetic fields are applied such that a small
of molecular dynamics applications [73, 76], where the
gradient is imposed on the resulting magnetic field.
reductions of CH dipole-dipole couplings due to fast
Depending on the geometry of the coils used for these
anisotropic dynamics were measured quantitatively. Its
PFGs, the gradients can adopt basically any geometry. The
signal modulation reads [65].
most common ones in high-resolution NMR are linear
gradients along the direction of the external field B0 (so-
SSS ~ Σ sin Nexc Φ (i) (0) sin Nre c Φ (i) (t1 )
i called “z-gradients”). Effectively, such PFGs introduce a
inhomogeneity into the B0 field and give rise to a well-
Πj cos N rec Φ (j) (t1 ) (15) defined dephasing of all transverse spin states, which can be
=i reversed by application of an inverse PFG. Based on these
14 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

dephasing and rephasing effects, PFGs are well established are relatively weak and need to be observed despite the
tools for the selection of coherence transfer pathways in presence of the strong 1H signal of long alkyl side chains
high-resolution NMR experiments [80, 81]. Their particular [92]. PFG-assisted suppression of this strong perturbing
advantage over pulse phase cycling schemes is that they signal is shown to be particularly useful for obtaining
develop their effect within a single transient of the pulse unambiguous results. Moreover, PFGs are also used in
sequence. Consequently, signal subtraction procedures over a inverse (1H) detected 1H-15N correlation experiments to
number of consecutive transients can be avoided, which remove strong unwanted 1H signal contributions [64, 65].
helps to reduce experiment time requirements and, moreover, Probing Molecular Orientation [93]
allows the dynamic range of the signal receiving unit to be
optimally adjusted to the incoming signal of interest. The recoupling schemes and PFGs approaches described
in the previous sections were concerned with providing
In solid-state NMR, however, PFGs are relatively rarely means to experimentally determine dipole-dipole couplings
used [82-85], because their usual application times are in the in a sensitive and selective fashion. Such couplings contain
order of a few milliseconds and, thus, relatively long as information on the structure and dynamics of molecules,
compared to transverse relaxation times of solids. Transverse because they depend on the distance of the two nuclei
spin states, which are required for PFGs to take effect, tend involved as well as on the orientation of the coupling vector
to be subject to strong spin-spin relaxation so that the signal relative to B0. In liquid-state NMR, these dependences are
largely disappears before a dephasing/rephasing cycle is also exploited in various ways, for example to derive
completed. Hence, PFGs need to be combined with distance constraints from proton-proton contacts or to infer
decoupling techniques that efficiently average the molecular conformations from residual dipole-dipole
interactions responsible for the relaxation mechanisms, i.e. couplings after partially aligning the molecules in a liquid-
predominantly dipole-dipole couplings in spin-1/2 systems. crystalline medium which adopts a preferred orientation
In heteronuclear 1H-X systems, conventional RF 1H along the B0 axis [22-24]. The latter approach has recently
decoupling pulses would serve this purpose, but due to the gained considerable importance for biological NMR
action of the field gradient the RF bandwidth has to be applications, and emphasises the relevance of orientation
correspondingly broad, which is usually beyond the measurements by NMR.
capabilities of available RF decoupling pulse schemes. In
addition, RF pulses and field gradient pulses would have to In solid-state NMR, such measurements are very well
be applied at the same time, which also makes high known, because the anisotropies of the interactions, in
demands on the NMR probe electronics; in particular, particular CSAs and 2H quadrupole couplings, allow the
coupling phenomena between the individual circuits need to orientation of molecular segments to be straightforwardly
be prevented. Magic-angle spinning (MAS), in contrast, detected by simple solid-state NMR spectra recorded on
does not give rise to such interference problems with PFGs, static samples [31]. The restriction to static samples means
and therefore appears as the method of choice. that the sites of interest need to be selectively labelled with
NMR-active isotopes, such as 13C or 2H, because an
Recently, PFGs have been made available to solid-state analysis of the data is only possible when a single or very
NMR by technically combining them with fast MAS few resonances contribute to the spectrum. To measure
equipment and experiments [78]. The required field gradients molecular orientation at multiple sites, the orientation
are provided by a simple pair of coils which have been built information needs to be correlated with a spectral dimension
into a standard fast-MAS probe. The selective dephasing and which provides chemical-shift resolution, and this is exactly
rephasing capabilities of PFGs are used to design how such experiments are performed in high-resolution
experimental techniques for 1H signal selection and NMR. An analogous strategy can be followed in solid-state
suppression in solids, which have originally been developed NMR when orientation measurements are performed under
for liquid-state NMR. In particular, WATERGATE [86-89]. (fast) MAS conditions, which can efficiently be
and DANTE [90, 91]. pulse sequences can be used under accomplished by using the orientation dependence of
MAS to selectively excite or suppress peaks in 1H solid- recoupling pulse sequences [93].
state spectra. These tools can readily be incorporated into
solid-state MAS NMR experiments, as has, for example, From Eqs. 6, 8a and 8b, it can be seen that the
been demonstrated for 1H-1H double-quantum NMR spectra recoupling efficiency depends on the orientation of the PAS
of supramolecular systems, where the 1H signals of interest in the rotor frame, namely on the angle β which represents

Fig. (7). (a) Pulse sequence for an inverse (1 H) detected 1 H-15 N correlation experiment in the solid state under fast MAS. (M2, M3) The
sequence consists of a TEDOR-type polarisation transfer, in the middle of which a t1 ’ dimension introduces rotor encoding, while a
spectral t1 dimension provides a 15 N spectrum with chemical-shift resolution (split-t1 approach). PFGs help with suppressing
unwanted 1 H signal. (b) Pair of field-gradient coils attached to the top and the bottom of a Bruker 2.5 mm fast-MAS stator in a
commercial 1 H-X double-resonance probehead with a standard bore diameter (54 mm).
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 15

the tilt between the rotor axis and the main axis of the magnetic quantum numbers ±1 are not affected by the 2H
anisotropic interaction. In detail, the ω R- and 2ω R- quadrupole coupling. Consequently, 2H DQ resonances do
modulated terms exhibit different orientation dependencies, not exhibit a quadrupolar lineshape. The same phenomenon
namely sin 2β and sin2 β, respectively. Obviously, from a constitutes the fundamental idea behind multiple-quantum
sin 2β dependence, the angle β can only be extracted (MQ) MAS NMR spectroscopy of half-integer quadrupolar
unambiguously over the interval β = [0°,45°], while the nuclei, which was initially proposed by F RYDMAN and
sin 2 β function is invertible over the interval β = [0°,90°]. HARWOOD [96], and has emerged as a powerful branch of
Hence, 2ω R-recoupling widens the range of distinguishable solid-state NMR spectroscopy with widely spread
molecular orientations by a factor of two. Moreover, in β- applications [97-103].
ranges where the angle sensitivity of sin 2β is relatively low, By combining the ideas of MQMAS and OMAS, in a
sin 2 β depends strongly on β and vice versa. In this way, the sense, a two-dimensional 2H NMR experiment can be
ranges with high β-sensitivity are complementary in the ω R- designed, which correlates isotropic 2H chemical shifts in
and 2ω R-modulated terms [93]. the indirect dimension with scaled pseudostatic 2H
In ref. [93] the effects of sample orientation have been lineshapes in the direct dimension. In this way, it is
investigated using highly oriented polyethylene fibres as a possible to separate multiple 2H resonance lines according to
model system, and the feasibility of extracting orientational the DQ chemical shifts of the involved 2H nuclei. In ref.
order parameters from sideband patterns of rotor-encoded [95] the theory of such two-dimensional 2H DQ OMAS
MAS experiments has been demonstrated. In the solid-state, experiments is described, and the principal features and
sample orientation phenomena are of particular interest for applicability is demonstrated using a sample of L-histidine
biological systems or for supramolecular structures, such as hydrochloride monohydrate, which was deuterated at four
discotic liquid crystals, since the performance of such positions by simple recrystallisation from D2O.
materials in optoelectronic devices strongly depends on the
degree of orientational order within the material [94]. In For 2H NMR spectroscopy, this DQ OMAS experiment
particular, an extruded fibre of a columnar discotic mesogen is particularly helpful because 2H virtually always needs to
[73]. has been studied by novel ω R- and 2ω R-recoupling be isotopically enriched in the samples. This in turn
methods, and the MAS NMR results have been compared to provides a practically perfect site selectivity of 2H NMR
X-ray diffraction studies [93]. investigations, as long as chemical synthesis allows the site
of interest to be deuterated in a fully selective fashion.
Scaling of Couplings by OMAS [95] Although, in chemical synthesis, the “art” of site-selective
Instead of using the different orientation dependences of deuteration has gained a remarkable degree of proficiency
ω R- and 2ω R-recoupling methods, orientation measurements [104, 105], the required synthetic efforts may still form an
can also be based directly on the evolution of the spin undesirable obstacle that calls for simplification. In this
system under an orientation dependent (i.e., anisotropic) context, a particular challenge is encountered for materials
interaction, provided that the anisotropies are not where simple deuteration procedures are no longer selective,
(completely) removed by an averaging technique. In the case but yield samples with multiple sites carrying 2H. This is,
of MAS, anisotropic interactions can straightforwardly be for example, always the case for acidic protons which can
allowed to partially “survive” the averaging process by conveniently be replaced by deuterons by exposing the
slightly misadjusting the magic angle. Under these off-MAS material to a deuterated solvent capable of exchanging its
(OMAS) conditions [31], anisotropic interactions are scaled deuterons with the solubilised material. Using deuterated
by 12 (3cos2 θ − 1) , which would be zero for θm = 54.7°. water, D2O, for example, such a simple exchange procedure
Technically, slight misadjustments in the range of a few places 2H at all sites of sufficient acidity, i.e.,
degrees can easily be realised using standard MAS predominantly NH n and OH positions in organic materials,
equipment. Since the effective scaling factor is then still while CHn sites are usually unaffected. 2H NMR
relatively small, this OMAS approach is basically restricted investigations of such multiply deuterated samples are
to strong anisotropic interactions. At this point, however, a considerably aggravated by the fact that the spectra consist of
contradiction is encountered, as residual anisotropic multiple superimposed lines which are, in general, difficult
interactions would of course lead again to significant line to separate by deconvolution procedures without perturbing
broadening and destroy the chemical-shift resolution. The the shapes of the individual resonance lines.
solution to this problem is to separate the measurement of
the (residual) anisotropic interaction from the chemical-shift 3. INTRODUCING SOLID-STATE NMR TECHNI-
information in two spectral dimensions. QUES INTO HIGH-RESOLUTION MAS NMR
A particularly intriguing way to realise this separation is
possible in the case of 2H NMR spectroscopy and the 2H When high-resolution NMR spectroscopy is performed
quadrupole coupling because of a fortunate combination of on systems in dilute solutions, the spectral resolution of the
two properties: Firstly, the 2H quadrupole coupling in solids resonance lines is primarily an issue with respect to the
is strong enough to allow all relevant information to be strength and the homogeneity of the static magnetic field.
extracted from a 2H NMR lineshape even if its spectral Aiming at maximum separation of sharp resonance lines,
width is considerably reduced under OMAS conditions. ongoing technical efforts are devoted to the increase of the
Secondly, the first-order quadrupole coupling affects all magnetic field (presently reaching about 1 GHz 1H
transitions and, thus, NMR frequencies of the spins except frequency) while possibly retaining its homogeneity (at a
for the maximum order transition. Therefore, the 2H double- level of < 0.1 Hz). As long as the systems under
quantum (DQ) transitions between the spin states of investigation can be isotropically dissolved in suitable
16 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

solvents, residual anisotropies of internal interactions do not The extreme situation of anisotropic interactions causing
play a significant role with respect to spectral resolution. severe line broadening and obscuring chemical shift
information is familiar from solid-state NMR and has been
In fact, liquid-crystalline media are increasingly used to
discussed in the previous Chapter. In partially mobile
introduce a small and directed anisotropy of the samples by
systems, the situation is certainly not as extreme as in
effectively aligning the molecules to some degree along the
solids, but when standard solution-state NMR experiments
external magnetic field [22-25]. Even under these conditions,
fail due to line broadening effects of residual anisotropic
the spectral resolution is practically not hampered. The
interactions, techniques from solid-state NMR appear to be
success of this relatively new approach and the significance
well suited to help overcome these problems. Following this
it gained over only a few years clearly indicates that systems
idea, MAS has successfully been combined with standard
with residual anisotropies, be they deliberately introduced or
solution-state NMR techniques, such as COSY [10], HMQC
be they inherently present, are about to become more and
[106, 107], HSQC [108] or NOESY [109], and the approach
more relevant in NMR research. Fields of applications
has been termed high-resolution (HR) MAS [26-28]. In the
include, among others, solid-phase synthesis and on-bead
majority of current HRMAS applications, however, MAS is
characterisation, tissue samples, or membrane proteins.
merely used to make the system behave more liquid-like and
Therefore, NMR methodology needs to make experiments
to allow standard solution-state experiments to be performed
and techniques available which are capable of delivering the
[110-112]. Considering the character of ”borderline” samples
desired pieces of information on such systems.

O
O
H6' O Si
PivO H2
H6
H4 O
PivO NH2
H5 OPiv
H3 H1

Fig. (8). 1 H NMR spectra of a small glycoside attached to swollen beads of polystyrene resin, as commonly used in solid-phase
synthesis. The spectral resolution is obscured mainly due to susceptibility effects which can be averaged by MAS.
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 17

that are neither isotropic solutions nor true solids, further typical for “inhomogeneous” line broadening, following the
progress can thus be expected when solid-state and solution- terminology introduced by MARICQ and WAUGH [35]. At
state NMR methods are combined more extensively. first sight, this appears a bit surprising from a solid-state
NMR point of view, because proton-proton dipole-dipole
In this work, the concept of recoupling, which is well
couplings are usually expected to play the dominating role
established in solid-state NMR, is introduced to HRMAS.
in line broadening. Their effect, however, would be of
In the HRMAS experiments performed so far, MAS was
homogeneous nature, which means that the lines should
used to efficiently remove anisotropic interactions
gradually narrow on increasing the MAS frequency. As this
throughout the experiment. The line-narrowing effect of
does not fit to the observations, the broadening must be of
MAS, however, is only required during the spectral
different origin. In this context, one needs to recall typical
dimensions, but not during the periods where the (residual)
effects encountered in high-resolution NMR and, in
interactions are essential as a driving force for informative
particular, in magnetic resonance imaging, where the
processes, such as exchange of magnetisation. By
heterogeneity of the sample is known to affect the achievable
introducing suitable recoupling techniques, this source of
resolution. In this respect, technical precautions are routinely
information becomes accessible and can be exploited for
taken in high-resolution NMR by using an optimised
characterising the systems under investigation more
sample geometry (long glass tubes) and, moreover, by
thoroughly.
slowly spinning the samples.
In the dipolar recoupling experiments presented below,
In HRMAS studies, the heterogeneous character of many
the anisotropic interactions are still efficiently averaged by
systems (e.g., mixtures of solvent and resin beads) gives rise
MAS during the t1 and t2 dimensions, thereby ensuring
to significant susceptibility gradients. The magnetic
spectral resolution, but the dipole-dipole couplings are active
susceptibility, χ, is a phenomenological factor relating the
during the exchange periods at (nearly) their “non-MAS”
magnetic field vector H0 with the magnetisation M acquired
strength. In addition, dipolar recoupling pulse sequences can
by the (nonferromagnetic) material: M = χH0. The magnetic
be designed such that only the dipolar interaction is re-
induction B0 (which, in NMR literature, is commonly,
introduced under MAS conditions, while unwanted
though in fact incorrectly, referred to as the “magnetic field”)
anisotropic interactions are suppressed. Moreover, the
is proportional to the sum of magnetic field and
possibility of tailoring average interaction Hamiltonians by
magnetisation where µ0 is the vacuum permeability. As long
RF pulse sequences also allows dipolar decoupling to be
as χ is a global parameter that is valid throughout the
accomplished and, in this way, an isotropic mixing
sample, it merely leads to a minor overall shift of B0.
Hamiltonian to be generated, which gives rise to exchange
However, when the quantity χ is considered as a scalar
processes purely driven by J-couplings. In the following, it
whose value is a local property χ(dV) of fractional sample
will be demonstrated how dipolar and J-driven exchange can
volumes dV, the resulting magnetisation is also a local
be distinguished and independently measured in two-
property. Consequently, each nuclear spin experiences local
dimensional NMR correlation experiments, corresponding to
dipolar fields arising from the local magnetisation of
the distinction of “through-space” and “through-bond”
surrounding fractional volumes. In analogy to the dipolar
contacts, by a combination of dipolar recoupling and
spin-spin coupling, the interaction can be viewed as a
decoupling schemes under HRMAS conditions.
dipole-dipole coupling between a spin and the local
magnetisation M(i) of fractional sample volumes dVi in the
3.1. The Quest for Spectral Resolution proximity ( of the spin. The corresponding Hamiltonian
Hx α ΣiM Zi) IZ is analogous to that of a heteronuclear
Considering systems with small residual anisotropies, dipole-dipole coupling where the second spin is replaced by
the effective magnetisation ΣiM Zi as experienced by the
()
the fundamental motivation for applying MAS in NMR
experiments is no doubt the lack of spectral resolution under spin I. [28].
static conditions. On the top of Fig. 8, a static 1H spectrum
is shown which is typically observed when samples from B0 = µ0 (H0 + M) = µ 0 (I + X ) H 0 , (16)
routine solid-phase synthesis are investigated. In such cases, Magic-Angle Spinning
the systems of interest, mostly peptides or glycopeptides,
are attached to swollen polymer resins. This greatly Based on this analogy, the effect of MAS on a sample
simplifies the synthetic procedures, but causes severe exhibiting susceptibility gradients can straightforwardly be
problems in the 1H NMR spectra, because they are understood: While in the static case the resonance lines are
dominated by the broad resonances of the resin, while the “inhomogeneously” broadened by the additional couplings
signals of the attached systems are totally obscured. As Hχ arising from local susceptibility differences, MAS
illustrated in Fig. 8, this problem can largely be overcome averages these interactions to zero and narrows the lines. In
by applying MAS. Moderate spinning frequencies of about principle, the situation is complicated by the fact that the
4 kHz usually suffice to remove essentially all line susceptibility χ is not a scalar quantity, but rather a tensor
broadening effects. due to its anisotropic properties. The problem arising from
the anisotropy is that H0 may no longer parallel B0 such that
Residual Anisotropies and Line Broadening the setting of the magic angle relative to the external field
Considering the MAS spectra at low spinning H0 is no longer correct with respect to B0. Thus, for
frequencies more closely, it becomes obvious that the broad appreciably anisotropic χ, MAS is effectively carried out
lines more or less instantaneously split into a bunch of sharp under slight “off-angle” conditions and cannot develop its
sidebands as soon as the sample is rotated. This behaviour is full averaging performance. This is, however, not the
18 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

dominating effect, as MAS is usually capable of averaging by LEVITT and coworkers [114, 115], have proven to
susceptibility gradients and, thus, provides satisfactory perform well under HRMAS conditions, and were therefore
resolution. used in the experiments to recouple homonuclear 1H-1H
dipole-dipole couplings.
Thus, with respect to line broadening, the residual
anisotropies of internal interactions play often only a minor Helical pulse sequences are based on the simultaneous
role in heterogeneous systems. The molecular dynamics, rotation of space and spin part through MAS and RF pulses,
though slightly restricted, is usually pronounced enough to respectively, and aim at creating an average Hamiltonian
ensure efficient pre-averaging and to leave only small consisting of selected tensor components Al,mT λ,µ by means
anisotropies, which can then be removed easily and of symmetry principles. The selection rules and the design
completely by MAS at low spinning frequencies. of appropriate sequences are described in detail in ref. [115].
Nevertheless, there are residual anisotropies in systems of To accomplish dipolar recoupling for 1H nuclei, a pulse
this kind, and the work described in the following aims to sequence is required which recouples the homonuclear
exploit their information content for determining molecular dipole-dipole coupling, while providing optimum
structure and dynamics. suppression of all other, potentially perturbing, interactions,
in particular chemical shift or, equivalently, off-resonance
effects. By tailoring the tensor component T 2,0 in the spin
3.2. The Creation of Coherent Spin States part, the properties of the “original” dipole-dipole coupling
can be retained (see Eq. 1). T 2,0 gives rise to zero-quantum
When MAS NMR experiments are performed on partially
coherences (ZQCs), involving “flip-flop” type transitions
immobilised systems, anisotropic interactions are averaged  ↓ ↔ ↓
↓ ↓
in pairs of coupled spins. It might appear
both by molecular dynamics and by MAS. The weak
natural to recouple this component, but it bears the
residual interactions, which are left over by these two
disadvantage that unwanted interactions are difficult to
averaging processes, merely give rise to relaxation effects.
suppress, because components T 2,0 exist for all interactions.
These can be understood and interpreted in a
A unique selection of the dipolar interaction is possible
phenomenological manner, but the direct information on
through its components T 2,±2. By recoupling these
individual couplings is lost. In order to restore this
components, dipolar double-quantum coherences (DQCs) are
information, the relaxation phenomena need to be replaced
generated between pairs of coupled spins, involving
by actual quantum-mechanical evolution processes which
transitions of the ”flop-flop” type  ↔ ↓↓ .
↓↓
directly and individually reflect the action of the couplings
on certain spins. This requires the interactions of interest to In principle, both ZQCs and DQCs can be used for
be amplified such that they emerge out of the “background dipolar exchange spectroscopy. Although for the DQ
noise” of fluctuating interactions of various kinds. process, in contrast to ZQCs, the energy of the spin system
is not conserved, the DQ approach is favourable in terms of
With respect to the experiments and systems discussed
suppressing unwanted interactions. In addition, as will be
here, interactions can only be amplified in terms of
discussed later, it imposes a quantum-chemical evolution on
preventing them from being reduced. While it is naturally
states of longitudinal magnetisation in NOESY-type
not possible to compensate for the pre-averaging effect of
experiments. The DQ-type of dipolar recoupling is realised
molecular motions by means of NMR techniques, it is
in the C7 21 pulse-sequence [114], which selects the terms
possible to prevent the additional averaging effect of MAS
A2,1T 2,2 and A2,-1 T 2,-2 of the dipolar Hamiltonian. For an ij
by recoupling pulse sequences. In this way, the spin system
spin pair the resulting average Hamiltonian read [114, 115]
experiences no longer weak residual interactions, but instead
where the coefficient ω D is given by.
recoupled interactions at their “original” strengths which
they would develop in the static case, i.e., in the absence of Dij is the dipolar coupling constant of the ij pair, and
MAS. This “non-MAS” strength can suffice to make the ω Rt0 is the rotor phase at the start of the pulse sequence. β
spin systems evolve in a quantum-mechanical manner such and γ denote the azimuthal and polar angle of the
that individual interactions become detectable and internuclear vector in the rotor frame. Apart from the
measurable. In the following section, this strategy will be orientation dependence and the phase factors, the average
discussed for the case of homonuclear dipole-dipole Hamiltonian of C721 contains an overall scaling factor of
couplings among protons. The same principles apply ~ 0.232 for the recoupled dipole-dipole coupling in
analogously to CSAs as well as to heteronuclear dipole- comparison to the static case [114].
dipole and quadrupole couplings.
HC7 = ωD T2,2 + ωD T2,-2 + 2πJ ij I(i) I(j) , (17)
Recoupling Schemes [113]
In systems with pronounced molecular mobilities,
anisotropic interactions are in general relatively weak, dipole-dipole J-coupling
typically on the order of a few Hertz. To ensure efficient 343(i + e iπ/14)
(18)
recoupling under these conditions, the pulse sequences have ωD = Dij sin(2β) e i(ωR t0-γ)
to be applied for durations of 100 ms and more, which is 520π 2
very long compared to typical timescales of solid-state “Through-Bond” and “Through-Space” Information
NMR. Thus, the pulse sequences need to be equipped with [113]
internal compensation mechanisms for the effects of other
interactions, experimental misadjustments and pulse It is important to note that, in addition to the dipolar
imperfections. In practice, helical pulse sequences, developed part, the average Hamiltonian in Eq. 17 also contains the
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 19

isotropic J-coupling between the spins i and j. The two can be restored and investigated at effectively ~ 20% of their
types of couplings differ in the information they convey, as “non-MAS” strengths. Apart from this “amplification”
J-couplings propagate through chemical bonds, while dipole- achievable by recoupling, there are important differences
dipole interactions act directly through space. In fact, J- between the Hamiltonians of C7 21-recoupled and (weak)
couplings are part of the average Hamiltonian not only for residual dipole-dipole couplings, which are obvious from
C7 21, but for all possible variants of helical pulse sequences. their spin parts:
This invariability of J-couplings opens up the possibility of
creating an average Hamiltonian which solely consists of J- HDC7 ~ T2,2 + T2,-2 ~ I (i) I(j) + 1 (i) I (j) (21a)
couplings, while all other interactions are suppressed due to offset (i) (j)
HD ~ 3I(i) (j) (i) (j)
Z IZ I I 2I Z IZ (21b)
their symmetry and transformation properties. Using helical
pulse sequences, the simplest scheme providing a pure J- The recoupled and residual interactions exhibit DQ and
coupling Hamiltonian is C931 [116, 117]. The average ZQ character, respectively. For residual couplings, in
Hamiltonian is simply given by which represents an addition, second averaging effects need to be taken into
isotropic mixing Hamiltonian. The coupling between the account, since the chemical-shift difference between the spins
spins i and j is completely independent of the molecular i and j often exceeds the residual coupling and truncates the
orientation and is also unaffected by resonance offsets, which dipolar Hamiltonian by suppressing the “flip-flop” exchange
would cause second averaging effects as soon as the term (see Eq. 21b). Dipolar recoupling, in contrast, prevents
frequency difference of the two resonances exceeds the J- such second-averaging effects, because the pulse sequence
coupling. In the context of isotropic mixing, a number of removes chemical-shift and resonance-offset contributions to
pulse sequences have been proposed in liquid-state NMR the average Hamiltonian.
[118, 119, some of which have recently been re-examined
and optimised with respect to their performance under Thus, considering an( )arbitrary state of longitudinal
HRMAS conditions [120, 121]. Here, the C931 sequence magnetisation Ρ 0 α a IZi b I(i) Z , and its quantum-
was chosen in order to keep the approaches for dipolar mechanical evolution under the different types of dipolar
recoupling (C721) and isotropic mixing (C931) as similar as interactions, three cases need to be distinguished: Firstly,
possible for combination purposes: The difference the “fully truncated” Hamiltonian HD α 2I(i) I(j)
Z Z , does not
Hamiltonian HC7 - HC9 = ω D . T 2.2 + ωD* T2.-2 is a pure lead to any evolution of the state ρ0, as is reflected by the
“dipolar” Hamiltonian, from which J-couplings have been commutation relation
removed. It cannot be produced as an average Hamiltonian of (i ) (i) (j)
a pulse sequence, but its effect on the spin system can be a I Z + b IZ(j) , 2IZ I Z = 0 (22)
approximated by the difference of the signals observed in a
C7 21 and a C931 recoupling experiment where τ rcpl denotes Secondly, under the “original” homonuclear dipolar
the recoupling time in the experiment. In the above Hamiltonian, HD α 3I(i) I(j) (i) (j)
Z Z - 1 I a state of longitudinal
approximation, mixed terms arising from the non-vanishing magnetisation evolves in terms of a ZQ exchange process,
commutator [HC7, HC9] 0 are neglected. Such terms provided that it is a non-equilibrium state with a = b ,
would only contribute to second and higher orders and have because the respective commutator reads
therefore no effect for two-spin systems. Even when multiple
spins are coupled, there is practically no effect for HC7 .τrepl (i ) (j) (i ) (j )
a IZ + b IZ , 3IZ I Z I(i)I(j ) =
<1 and HC9 .τrepl <1. Thus, by combining a C721 and a (j )
C9 31 recoupling experiment, it is possible to record NMR 2 (a b) (I (i) I I(i) I(j) ) (23)
spectra which contain information on either dipole-dipole Thirdly, the recoupled dipolar DQ Hamiltonian HC7 D
couplings or J-couplings [113]. In this way, “through-bond” generates DQ coherences out of longitudinal magnetisation,
contacts can be distinguished from “through-space” contacts as long as a = -b . These differences between the three cases
between spins. have particular consequences for NOESY-type experiments
H C9 = 2πJ i j .I (i) I (j) (19) (see below), where during a mixing time exchange processes
occur between dipolar-coupled spins that occupy states of
exp i(H C7 HC 9 )τ rc pl ρ 0 exp i(H C7 H C9 )τ
rc pl
longitudinal magnetisation. For residual dipole-dipole
couplings represented by a “fully truncated” Hamiltonian,
≈ ρ 0 + exp iH τrcpl ρ 0 exp τrc pl
C7 C7
iH there is no quantum-mechanical evolution of the spin
system. Consequently, exchange processes are solely based
on diffusion of magnetisation, which occurs on much longer
C712 -experiment time scales than quantum-chemical evolution. Such diffusive
processes are described in a phenomenological manner [10,
exp iHC9 τrcpl ρ 0 exp iHC9 τ rcpl 122] and are therefore difficult to quantify in terms of
individual two-spin couplings. In particular, the knowledge
of motional correlation times is required in order to
C912 -experiment determine dipole-dipole couplings from conventional
(20) NOESY data (see below).
Recoupled and Residual Interactions
By applying the C721 pulse sequence, MAS experiments a I (i)
Z
+ b I (j)
Z
,T 2.+2 = 2(a + b) T2,+2 (24)
are no longer restricted to the observation of very weak
residual dipolar interactions, but the dipole-dipole couplings
20 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

If the truncation of the dipolar Hamiltonian is avoided by well as 2H quadrupole couplings can be determined
appropriate recoupling, a quantum-mechanical evolution of selectively and accurately by different recoupling techniques.
longitudinal spin states is re-introduced. For ZQ- and DQ- Recoupled Exchange [113]
type Hamiltonians, this evolution is proportional to the
degree of non-equilibrium (a – b in Eq. 23) and equilibrium An experiment frequently used in high-resolution NMR
(a + b in Eq. 24), respectively. Conversely, the sum is two-dimensional 1H-1H NOESY (“Nuclear Overhauser
magnetisation Σ = I(i)Z
I (j)
Z and the difference magnetisation Enhancement SpectroscopY”) [109], the pulse sequence of
∆ = I Z I Z are conserved under ZQ- and DQ-type
(i) (j)
which is depicted in Fig. 9a. In the spectra, distinguishable
evolution, respectively. species of protons are correlated by signals (“cross peaks”)
which reflect a transfer of magnetisation from one species to
The latter property also opens up a possibility to the other. This transfer is an incoherent cross-relaxation
distinguish J-couplings from dipole-dipole couplings. The effect which is driven by fluctuating residual dipole-dipole
two types of couplings provide complementary pieces of couplings between the two spins involved. Using a
information about the arrangement of two spins, namely phenomenological model, the signal intensity can be
their molecular connectivity and their spatial proximity. In described by an exponential relation of the form [10, 122]
general, a connection via chemical bonds which causes a where the rate constants RC and RL stand for the cross-
detectable J-coupling comes along with a certain degree of relaxation and the “leakage” rate, respectively, and τ mix is
spatial proximity, but the precise measurement of the spin- the mixing time during which the transfer processes occur.
spin distance allows, for example, the molecular While the former contains the coupling between the two
spins ij of interest, RC = 1/ 5 D2ij τ c ,the latter represents the
conformation to be elucidated. Furthermore, the orientation
dependence of the dipole-dipole coupling can, in principle, interaction of the pair with further spins. However, the cross-
be used for determining the orientation as well as, in the correlation rate, RC, depends not only on the coupling Dij,
case of mobile systems, the local mobility and dynamics of but also on the motional correlation time τ C, which is
molecular segments. In standard solid- and liquid-state commonly used to distinguish between the limits of fast and
NMR, dipole-dipole couplings and J-couplings do not slow motion with respect to the time scale of the Larmor
interfere and there is no need to unravel them, because one of frequency, ω 0. For the samples investigated here, the
the two interactions clearly predominates. In partially mobile validity of the slow motion limit (also called “spin diffusion
systems, however, the motional averaging is incomplete, limit”), i.e., τ c >> 2π . ω 0-1 = 10 ns, can be safely
such that the two types of couplings can effectively be of assumed, but the precise value of τ C is not known, such that
comparable strength, and means of distinction become the evaluation of the NOESY data alone yields relative
valuable. dipole-dipole coupling constants only.
In conclusion, the dipolar recoupling procedures outlined
e-RC τ mix ) e-RC τ mix ,
above take advantage of (25)
S(t) ~ (1
• the amplification, i.e. recovery, of dipole-dipole Instead of relying on such cross-relaxation phenomena,
couplings under MAS, the homonuclear 1H-1H dipole-dipole coupling can be
• the absence of second averaging, recoupled and, effectively, be amplified under MAS
conditions, as discussed above. For this purpose, a dipolar
• the presence of quantum-mechanical evolutions recoupling sequence needs to be applied during the mixing
instead of relaxation processes, and time (see (Fig. 9b). In this way, the incoherent cross-
• the conservation of ∆ instead of Σ, when DQ-type relaxation processes are replaced by coherent transfer
recoupling is performed, which provides a means to processes via dipolar DQCs, and the signal intensities can be
distinguish homonuclear dipole-dipole from J- expressed as where the “dipolar frequency” ω D is given by
couplings. Eq. 18 when the C721 sequence is used. In this way,
residual dipole-dipole couplings Dij can be determined from
These features make the C721 pulse sequence an attractive experimental signal intensities S(τ mix ) observed for
candidate for replacing the diffusive exchange process in individual spin pairs ij. The 1H HRMAS spectra resulting
MAS NOESY experiments. The above considerations from the simple NOESY experiment and the “C7-recoupled”
focused on homonuclear dipole-dipole couplings between experiment are shown in Figs. 10a and 10b, respectively,
protons, but they apply analogously to all interactions and two major differences are evident: Firstly, the mixing
recoupled by appropriate pulse sequences, in particular to times required for efficient transfer processes are shorter by a
quadrupole and heteronuclear dipole-dipole couplings. factor of ~ 25 under dipolar recoupling conditions, because
the couplings are amplified by preventing the averaging
3.3 The Measurement of Couplings effect of MAS. Secondly, the “C7-recoupled” exchange
signals are (partially) negative in the spectra. This proves
After discussing the motivation for and the principal that the transfer occurs in a coherent fashion via DQCs,
concepts of recoupling approaches under HRMAS because during this process the difference (and not the sum)
conditions, it is time to turn to measurements of couplings magnetisation of the respective spin pair ij is conserved (see
which have been performed by recoupling experiments. In above), which renders the exchange signals negative.
the following, it shall be demonstrated how homonuclear
1H-1H and heteronuclear 1H-13C dipole-dipole couplings as S (τ mix ) ~ sin2 ( 1 2 ωD τ mi x) β, γ (26)
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 21

In the exchange experiments discussed so far, DQCs of ω i) and (ω j, ω j), or a single signal at (ω i, ωi), if the
spin pairs ij serve as transition states for i j magnetisation chemical shifts of i and j are identical. As a consequence of
transfer processes. Instead of merely using DQCs as this, in exchange spectra, signals tend to overlap on the
transition states, they can also be spectroscopically detected diagonal, and exchange between like spins is difficult to
in a (indirect) DQ dimension that is inserted into the mixing observe, while the respective signals are separately observed
time and replaces the t1 dimension of the exchange in DQ spectra. In fact, the appearance of DQ signals arising
experiment (see schemes in Figs. 9b and 9c). The resulting from pairs of like spins is a noteworthy feature of dipolar
dipolar DQ-SQ correlation experiment is usually referred to DQ spectra. In the case of J-mediated correlation spectra (i.e.
as dipolar DQ spectroscopy, which has been established in INADEQUATE), such signals are not observable because
solid-state MAS NMR [21, 68]. It bears analogy to the they rely on the anisotropy of the interaction.
INADEQUATE [123, 124] experiment known from liquid- The DQ signal intensity depends on the dipole-dipole
state NMR, which in contrast makes use of J-couplings for coupling Dij according to Eq. 26, but experimentally, the
the generation of DQCs. From a two-dimensional dipolar oscillations are usually obscured by an exponential decay.
DQ spectrum, pairs of coupled spins can be directly This effect is known from solid-state 1H-1H DQ experiments
identified. Pairs of unlike spins, ij, give rise to two signals [21]. In the limit of short recoupling times, i.e.
placed at (ω i + ω j, ω i) and (ω i + ω j, ω j), while pairs of like Dijτ exc/rec/2π<1, the sine dependence in Eq. 26 may be
spins, ii, are observed on the DQ-SQ “diagonal” at (2ω i, approximated by the leading linear term, which yields
ω i). In an exchange experiment, in contrast, an ij correlation expressions of the form SD Q ~ ω D2 β.γ τ ex
2
c/rec and SDQ ~ rij
-
gives rise to four signals, placed at (ω i, ωi), (ω i, ωj), (ω j,

Fig. (9). Experimental schemes of conventional NOESY-type (a) and recoupled dipolar (b) exchange experiments. Instead of merely
serving as transition states, the dipolar DQCs can also be detected spectroscopically (c).

Fig. (10). Two-dimensional 1 H-1 H HRMAS spectra resulting from the experiments and pulse sequences depicted in Fig. 9: (a)
Conventional NOESY-type exchange, (b) “C7-recoupled” dipolar exchange, (c) DQ spectroscopy. Dotted contour lines indicate
negative signals. The sample under investigation, together with the assignment of the 1 H resonance lines, is given in Fig. 8.
22 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

6 (fora given recoupling time). Thus, the signals of dipolar initially developed by LEVITT and G RIFFIN, [126, 127] and
DQ spectra provide direct insight into the (average) distances belongs to the routine experiments in solid-state MAS
between distinct nuclei. Experimentally, distance ranges of NMR. The recoupling effect of RR is restricted to dipole-
up to rij ≤ 4 … 5 Å have found to be accessible [21]. dipole couplings between pairs of spins whose chemical-
shift difference exactly matches the applied MAS frequency
In the recoupled exchange experiments described so far,
(or a small integer multiple of it). The bandwidth of RR
effects potentially arising from J-couplings have been
depends on the ZQ-relaxation properties of the system [127-
neglected. In DQ spectroscopy, there are indeed no such
129], and was found to be around 10 Hz for proton-proton
effects, because only dipole-dipole couplings can give rise to
couplings under HRMAS conditions [125].
DQCs, while J-couplings cannot due to the ZQ-character of
their Hamiltonian (see Eq. 19). In exchange experiments, At the start of a RR exchange experiment, a state of
however, J-couplings may give rise to signals, which can maximum difference magnetisation, ρ0 α I(i) Z I(j)
Z , is
also been seen from the spectrum in Fig. 10b. The cross prepared by inverting the state of one of the two spins.
peaks are not purely negative, as they would be for recoupled Consecutively, the spins are allowed to exchange
dipolar exchange, but exhibit positive contributions from J- magnetisation during a mixing time under RR conditions,
couplings between the respective nuclei. In this way, a which ideally leads to a magnetisation difference of zero.
distinction of the effects is possible in a qualitative manner, The difference is recorded as a function of the mixing time,
but there is need for a quantitative separation, which can be and analysed by fitting calculated curves (see Fig. 12a).
accomplished by the combination of C721 recoupling and From the initial slope of the curve, the effective dipole-
C9 31 decoupling, as discussed in the previous section. (Fig. dipole coupling Dij between the spins can then be
11) shows the two-dimensional 1H-1H spectra observed for determined selectively. The RR experiment has been
C7 21-recoupling and C931-decoupling, as well as the “C7- introduced to HRMAS conditions, and 1H-1H dipole-dipole
C9” difference spectrum. The latter contains pure dipolar couplings as low as 0.3 Hz could be measured between two
(“through-space”) information and, correspondingly, only methyl groups [125]. The selectivity of the RR approach
negative cross peaks are observed. originally relies on the resolution of the 1H resonance lines
in the spectrum, but it can be significantly enhanced by
Rotational Resonance [125]
selective isotope enrichment if necessary. For example, 13C-
The C721 pulse sequence used above provides broadband labelled methyl groups can be used, whose 1H signals can be
recoupling of dipole-dipole couplings between all types of selected by means of a conventional high-resolution 1H-13C
protons in a system, while the selection of spin pairs ij HSQC/HMQC step [10, 29] before the RR experiment. In
relies fully on the resolution of the resonance lines in the this way, liquid-state (HSQC/HMQC) and solid-state (RR)
two-dimensional spectra. However, if spectral resolution is NMR methods are directly combined in a single HRMAS
the only means to separate information, it is not possible to experiment [125].
distinguish whether a transfer i ←
← ← → j occurs really directly Rotor Encoding and Sideband Patterns [125]
or via a third spin i ← k ← j . The latter effect is well
known as “relayed transfer”, and usually prevents the Under HRMAS conditions, the 1H-1H dipole-dipole
determination of weak couplings Dij in the presence of coupling determined by the RR experiment usually contains
stronger ones, Dik and Dkj. To overcome this problem, two pieces of information. Firstly, the couplings depend on
recoupling schemes are needed which selectively restore a the internuclear distances, rij; secondly, the couplings are
single type of coupling, while leaving all others unaffected. reduced by molecular motions that occur on timescales
A particularly efficient and robust method for selective below microseconds. In order to separate the structural and
dipolar recoupling is rotational resonance (RR), which was dynamic aspects by means of dipolar recoupling

Fig. (11). Two-dimensional 1 H-1 H HRMAS exchange spectra resulting from (a) C72 1 -recoupling, (b) C93 1 -decoupling and (c) the
difference of the two. The C93 1 spectrum contains pure “through-bond” information, because the exchange occurs only via J-
couplings. The “C7-C9” spectrum, in contrast, contains pure dipolar (“through-space”) information, because practically all
contributions from J-couplings are removed by the subtraction.
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 23

experiments, measurements need to be performed where one only a small residual anisotropy is left. Nevertheless, dipolar
of the two parameters (i.e., either the distance or the motion) recoupling approaches are still feasible.
is known. Obviously, this criterion is met for dipole-dipole Using the reduction factors S, the motional pre-averaging
couplings of chemically-bonded 1H-13C entities, as their of the homonuclear 1H-1H dipole-dipole couplings
(bond) distance is known. Consequently, such 1H-13C determined by RR experiments (see above) can be taken into
dipole-dipole couplings are only affected by motions of the account and removed from the coupling constants.
molecular segment they belong to. Depending on the molecular structure, it might be necessary
Experimentally, various pulse sequences are available to combine the reduction factors of different molecular
from solid-state NMR, which recouple heteronuclear dipole- segments in order to appropriately model the dynamics of
dipole couplings in the presence of MAS [51,52, 56]. For the molecule [125]. The “corrected” coupling constants,
HRMAS applications, the rotor-encoded REDOR Dij/S, so obtained depend basically only on the effective
(REREDOR) scheme has proven to be particularly useful, as distance between the protons i and j. In this way,
it combines technical robustness with a sensitive means of information on structure and dynamics can be separated from
detection, namely rotor-encoded sideband patterns (see each other, and effective (i.e., averaged) distances of up to
Chapter 2.3). Initially, REREDOR experiments have been 8 Å could be measured [125].
developed in this work as solid-state applications for fast Under HRMAS conditions, the local or segmental
MAS [65], but they can equally be applied under HRMAS dynamics of a molecule can be determined not only by
conditions. Fig. 12b shows three HRMAS REREDOR recoupling heteronuclear 1H-13C dipole-dipole couplings,
sideband patterns from which 1H-13C dipole-dipole but also by recoupling 2H quadrupole couplings. For this
couplings as low as 10 Hz can be determined independently purpose, a homonuclear recoupling scheme has to be
for the three methyl groups [125]. The ratio of these applied, which means that the REDOR-based pulse
measured couplings to the (hypothetical) value of a rigid sequences for 1H-13C recoupling need to be replaced by, for
group can be interpreted as a reduction factor S, which example, the back-to-back pulse sequence [21, 59]. The latter
represents the motional pre-averaging in terms of the residual recouples homonuclear dipole-dipole couplings as well as
anisotropy of the respective molecular segment. In the (weak) quadrupole couplings, because a homonuclear spin
tetrapeptide studied in ref. [125] these factors were of the system consisting of two dipolar-coupled spin-1/2 nuclei is
order of S = 2. 10-3 , which shows that the systems under formally identical to a quadrupolar spin-1 system. For such
investigation move rapidly and almost isotropically, and 2H NMR studies, the sample needs to be deuterated at the

Fig. (12). (a) Rotational-resonance decay curves observed for the two 1 H-1 H methyl-methyl couplings indicated above in the
tetrapeptide structure. (b) Sideband patterns of the three methyl groups, observed in a 1 H-13 C rotor-encoded REDOR experiment. The
respective dipole-dipole couplings obtained from the experiments are given in the figure. For the 1 H-13 C couplings, reduction factors
S are also given, which reflect the motion of the respective methyl group. To derive these, a 1 H-13 C coupling of D = 6.9 kHz was
assumed for a rigid methyl group (that only rotates rapidly about its threefold symmetry axis).
24 Current Analytical Chemistry, 2005, Vol. 1, No. 1 Ingo Schnell

sites of interest. Using the tripeptide depicted in (Fig. 13) as order from one aspartic-acid sidechain end to the other is
an example [125], deuterated methyl and methylene groups reflected by the ratio of the respective parameters, i.e.,
have been introduced by esterifying the acid functionalities SMe /SEt ≈ 0.8. This means that the peptide as a whole is
with fully deuterated methanol and ethanol. The resulting highly mobile (with S = 2 . 10-3 ) due to the flexibility of the
tripeptide sample is depicted in Fig. 13, together with linker by which it is attached to the polymer resin.
spinning sideband patterns which are generated by rotor- Internally, however, the peptide adopts a relatively immobile
encoding the recoupled quadrupole couplings. The 2H and stable structure (with S ≈ 0.8). It is therefore reasonable
chemical-shift resolution (see also ref. [95] suffices to to refer to a structure of the peptide molecule with noticeable
separate all three 2H resonances and to analyse the sideband internal dynamics and a pronounced overall motion [125].
patterns individually Fig. 13b. By comparing the
expeimental patterns with calculated ones (Fig. 13a), the
quadrupole couplings are determined, which can then be 4. CONCLUSIONS AND OUTLOOK
converted into motional reduction factors S by relating the
measured couplings to the 2H coupling of a rigid methyl After the fields of liquid- and solid-state NMR
group (δQ/2π = 40 kHz, where “rigid” means that each spectroscopy have developed largely independently of each
group only rotates about its threefold symmetry axis other over the past 50 years, materials of current interest,
[31, 129]. particularly in biological NMR, call for new approaches
which combine and merge spectroscopic techniques from
When the segmental dynamics has been determined by either side. In this respect, recent technical progress opens up
means of such parameters S at several sites of the same a number of possibilities to promote NMR methodology
molecule, it is further possible to characterise the internal into this direction, for example by employing high-speed
dynamics, or residual order, within the molecule. In the tri- MAS in solid-state NMR [21] and by performing high-
and tetrapeptides (Figs. 12 and 13), for example, the residual resolution NMR under moderate MAS conditions [28].

CD3 CD2
O
O

O O Si
H
N O
O N N
H H H
O O N
O
O O
O
CD3

Fig. (13). Molecular dynamics of the deuterated methyl and methylene groups in the given tripeptide structure, as determined by
sideband patterns resulting from the rotor-encoding of recoupled 2 H quadrupole couplings under HRMAS conditions. (a) Calculated
and (b) experimental patterns with different coupling constants (CQ = δ Q/2 π).
Merging Concepts from Liquid-State and Solid-State NMR Spectroscopy Current Analytical Chemistry, 2005, Vol. 1, No. 1 25

Based on this background, this review intended to provide Under HRMAS conditions, diffusive processes of 1H-1H
an overview of new NMR methods and experiments which magnetisation transfer can, for example, be replaced by
have been developed by merging concepts from liquid-state specific 1H-1H correlations via dipolar double-quantum
and solid-state NMR. coherences, when a suitable recoupling pulse sequence is
applied during the mixing period of 1H-1H NOESY
In solid-state NMR, a wealth of new and versatile
experiments. [113] Highly selective dipolar recoupling is
experiments can be designed by using coherent spin-states
provided by rotational resonance techniques, [127] which
and coherent transfer mechanisms. These concepts form the
enabled the selective determination of 1H-1H dipole-dipole
basis for virtually all multidimensional liquid-state NMR
couplings as weak as 0.3 Hz between two methyl groups of
experiments [11, 13, 29], and the principal ideas can be
an oligopeptide attached to a polymer resin [125].
adapted to solid-state conditions when (i) fast MAS ensures
Heteronuclear dipolar recoupling, e.g. 1H-13C, allows for the
efficient averaging of anisotropic interactions, and (ii)
measurement of residual dipole-dipole couplings of
recoupling schemes selectively re-introduce the interactions
molecular segments, which directly reflects the dynamics of
of interest, i.e., predominantly dipole-dipole couplings
these segments. The same information is, in principle, also
which basically serve as a substitute for the much weaker J-
accessible from residual 2H quadrupole couplings which can
couplings. In this way, a wealth of experimental schemes
also be measured by recoupling approaches under HRMAS
can be transferred from liquid-state to solid-state NMR
conditions. Notably, all these recoupling experiments
[63-65, 78, 95], including two important and well-
provide the coupling information in combination with
established techniques, namely inverse (i.e., 1H) detection
chemical-shift resolution, i.e., in a site-selective fashion.
schemes for sensitivity enhancement [63-65] and pulsed
field-gradients (PFGs) for signal selection and suppression Overall, MAS NMR experiments are attractive tools for
[78]. studying molecular structure and dynamics in solids as well
as in “borderline” materials which are neither rigid solids nor
Due to the anisotropy inherent to solid-state interactions,
isotropic solutions or liquids. The information on structure
recoupling schemes can be performed under MAS in such a
and dynamics is contained in anisotropic interactions, and
way that the recoupled interaction becomes “rotor-encoded”,
can therefore be obtained in a highly selective and accurate
which gives rise to characteristic sideband patterns in the
manner from recoupling experiments which involve the
spectra [65]. These patterns provide a very sensitive means
generation of coherent spin states. These strategies apply
to measure the underlying coupling, and can, in a sense, be
equally well to solid-state and high-resolution NMR, and
viewed as the solid-state MAS analogues to the typical line
the differences between these two traditional fields of NMR
splittings observed in high-resolution NMR spectra.
spectroscopy come down to a few technical details
Correspondingly, the sideband patterns can readily serve as a
concerning the fine adjustment of the experiments. As a
measure for molecular structure [65, 66, 130, 131-133],
consequence of this, there is hardly a reason to keep the
dynamics [134, 135], and orientation phenomena [93].
separation of solid- and liquid-state NMR. In fact, there is
Furthermore, it is possible to re-introduce and to detect weak
need to promote their “re-unification” in order to be able to
residual interactions in solid-state NMR by performing
investigate the vast field of materials which combine
MAS under “off-angle” conditions (so-called OMAS). This
properties of solids and liquids, but do not solely belong to
concept is particularly suited for 2H quadrupole interactions,
either side.
which can conveniently be determined by 2H double-
quantum OMAS spectroscopy in a site-selective fashion
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Received:26 August, 2004 Accepted: 15 September, 2004