Magic angle spinning NMR and 1H-31P heteronuclear statistical total correlation spectroscopy of intact human gut biopsies

Previously we have demonstrated the use of 1H magic angle spinning (MAS) NMR spectroscopy for the topographical variations in functional metabolic signatures of intact human intestinal biopsy samples. Here we have analyzed a series of MAS 1H NMR spectra (spin-echo, one-dimensional, and diffusion-edited) and 31P-{1H} spectra and focused on analyzing the enhancement of information recovery by use of the statistical total correlation spectroscopy (STOCSY) method. We have applied a heterospectroscopic cross-examination performed on the same samples and between 1H and 31P-{1H} spectra (heteronuclear STOCSY) to recover latent metabolic information. We show that heterospectroscopic correlation can give new information on the molecular compartmentation of metabolites in intact tissues, including the statistical "isolation" of a phospholipid/triglyceride vesicle pool in intact tissue. The application of 31P-1H HET-STOCSY allowed the cross-assignment of major 31P signals to their equivalent 1H NMR spectra, e.g., for phosphorylcholine and phosphorylethanolamine. We also show pathway correlations, e.g., the ascorbate-glutathione pathway, in the STOCSY analysis of intact tissue spectra. These 31P-1H HET-STOCSY spectra also showed different topographical regions, particular for minor signals in different tissue microenvironments. This approach could be extended to allow the detection of altered distributions within metabolic subcompartments as well as conventional metabonomics concentration-based diagnostics.     

Heteronuclear 19F-1H statistical total correlation spectroscopy as a tool in drug metabolism: study of flucloxacillin biotransformation

We present a novel application of the heteronuclear statistical total correlation spectroscopy (HET-STOCSY) approach utilizing statistical correlation between one-dimensional 19F/1H NMR spectroscopic data sets collected in parallel to study drug metabolism. Parallel one-dimensional (1D) 800 MHz 1H and 753 MHz 19F{1H} spectra (n = 21) were obtained on urine samples collected from volunteers (n = 6) at various intervals up to 24 h after oral dosing with 500 mg of flucloxacillin. A variety of statistical relationships between and within the spectroscopic datasets were explored without significant loss of the typically high 1D spectral resolution, generating 1H-1H STOCSY plots, and novel 19F-1H HET-STOCSY, 19F-19F STOCSY, and 19F-edited 1H-1H STOCSY (X-STOCSY) spectroscopic maps, with a resolution of approximately 0.8 Hz/pt for both nuclei. The efficient statistical editing provided by these methods readily allowed the collection of drug metabolic data and assisted structure elucidation. This approach is of general applicability for studying the metabolism of other fluorine-containing drugs, including important anticancer agents such as 5-fluorouracil and flutamide, and is extendable to any drug metabolism study where there is a spin-active X-nucleus (e.g., 13C, 15N, 31P) label present.     

"Add to subtract": a simple method to remove complex background signals from the 1H nuclear magnetic resonance spectra of mixtures

Because of its highly reproducible and quantitative nature and minimal requirements for sample preparation or separation, (1)H nuclear magnetic resonance (NMR) spectroscopy is widely used for profiling small-molecule metabolites in biofluids. However (1)H NMR spectra contain many overlapped peaks. In particular, blood serum/plasma and diabetic urine samples contain high concentrations of glucose, which produce strong peaks between 3.2 ppm and 4.0 ppm. Signals from most metabolites in this region are overwhelmed by the glucose background signals and become invisible. We propose a simple "Add to Subtract" background subtraction method and show that it can reduce the glucose signals by 98% to allow retrieval of the hidden information. This procedure includes adding a small drop of concentrated glucose solution to the sample in the NMR tube, mixing, waiting for an equilibration time, and acquisition of a second spectrum. The glucose-free spectra are then generated by spectral subtraction using Bruker Topspin software. Subsequent multivariate statistical analysis can then be used to identify biomarker candidate signals for distinguishing different types of biological samples. The principle of this approach is generally applicable for all quantitative spectral data and should find utility in a variety of NMR-based mixture analyses as well as in metabolite profiling.     

Statistical total correlation spectroscopy: an exploratory approach for latent biomarker identification from metabolic 1H NMR data sets

We describe here the implementation of the statistical total correlation spectroscopy (STOCSY) analysis method for aiding the identification of potential biomarker molecules in metabonomic studies based on NMR spectroscopic data. STOCSY takes advantage of the multicollinearity of the intensity variables in a set of spectra (in this case 1H NMR spectra) to generate a pseudo-two-dimensional NMR spectrum that displays the correlation among the intensities of the various peaks across the whole sample. This method is not limited to the usual connectivities that are deducible from more standard two-dimensional NMR spectroscopic methods, such as TOCSY. Moreover, two or more molecules involved in the same pathway can also present high intermolecular correlations because of biological covariance or can even be anticorrelated. This combination of STOCSY with supervised pattern recognition and particularly orthogonal projection on latent structure-discriminant analysis (O-PLS-DA) offers a new powerful framework for analysis of metabonomic data. In a first step O-PLS-DA extracts the part of NMR spectra related to discrimination. This information is then cross-combined with the STOCSY results to help identify the molecules responsible for the metabolic variation. To illustrate the applicability of the method, it has been applied to 1H NMR spectra of urine from a metabonomic study of a model of insulin resistance based on the administration of a carbohydrate diet to three different mice strains (C57BL/6Oxjr, BALB/cOxjr, and 129S6/SvEvOxjr) in which a series of metabolites of biological importance can be conclusively assigned and identified by use of the STOCSY approach.     

Statistical correlation and projection methods for improved information recovery from diffusion-edited NMR spectra of biological samples

Although NMR spectroscopic techniques coupled with multivariate statistics can yield much useful information for classifying biological samples based on metabolic profiles, biomarker identification remains a time-consuming and complex procedure involving separation methods, two-dimensional NMR, and other spectroscopic tools. We present a new approach to aid complex biomixture analysis that combines diffusion ordered (DO) NMR spectroscopy with statistical total correlation spectroscopy (STOCSY) and demonstrate its application in the characterization of urinary biomarkers and enhanced information recovery from plasma NMR spectra. This method relies on calculation and display of the covariance of signal intensities from the various nuclei on the same molecule across a series of spectra collected under different pulsed field gradient conditions that differentially attenuate the signal intensities according to translational molecular diffusion rates. We term this statistical diffusion-ordered spectroscopy (S-DOSY). We also have developed a new visualization tool in which the apparent diffusion coefficients from DO spectra are projected onto a 1D NMR spectrum (diffusion-ordered projection spectroscopy, DOPY). Both methods either alone or in combination have the potential for general applications to any complex mixture analysis where the sample contains compounds with a range of diffusion coefficients.     

Trace level detection of compounds related to the chemical weapons convention by 1H-detected 13C NMR spectroscopy executed with a sensitivity-enhanced, cryogenic probehead

Two-dimensional 1H-13C HSQC (heteronuclear single quantum correlation) and fast-HMQC (heteronuclear multiple quantum correlation) pulse sequences were implemented using a sensitivity-enhanced, cryogenic probehead for detecting compounds relevant to the Chemical Weapons Convention present in complex mixtures. The resulting methods demonstrated exceptional sensitivity for detecting the analytes at trace level concentrations. 1H-13C correlations of target analytes at < or = 25 microg/mL were easily detected in a sample where the 1H solvent signal was approximately 58,000-fold more intense than the analyte 1H signals. The problem of overlapping signals typically observed in conventional 1H spectroscopy was essentially eliminated, while 1H and 13C chemical shift information could be derived quickly and simultaneously from the resulting spectra. The fast-HMQC pulse sequences generated magnitude mode spectra suitable for detailed analysis in approximately 4.5 h and can be used in experiments to efficiently screen a large number of samples. The HSQC pulse sequences, on the other hand, required roughly twice the data acquisition time to produce suitable spectra. These spectra, however, were phase-sensitive, contained considerably more resolution in both dimensions, and proved to be superior for detecting analyte 1H-13C correlations. Furthermore, a HSQC spectrum collected with a multiplicity-edited pulse sequence provided additional structural information valuable for identifying target analytes. The HSQC pulse sequences are ideal for collecting high-quality data sets with overnight acquisitions and logically follow the use of fast-HMQC pulse sequences to rapidly screen samples for potential target analytes. Use of the pulse sequences considerably improves the performance of NMR spectroscopy as a complimentary technique for the screening, identification, and validation of chemical warfare agents and other small-molecule analytes present in complex mixtures and environmental samples.     

Optimization of human plasma 1H NMR spectroscopic data processing for high-throughput metabolic phenotyping studies and detection of insulin resistance related to type 2 diabetes

Optimizing NMR experimental parameters for high-throughput metabolic phenotyping requires careful examination of the total biochemical information obtainable from (1)H NMR data, which includes concentration and molecular dynamics information. Here we have applied two different types of mathematical transformation (calculation of the first derivative of the NMR spectrum and Gaussian shaping of the free-induction decay) to attenuate broad spectral features from macromolecules and enhance the signals of small molecules. By application of chemometric methods such as principal component analysis (PCA), orthogonal projections to latent structures discriminant analysis (O-PLS-DA) and statistical spectroscopic tools such as statistical total correlation spectroscopy (STOCSY), we show that these methods successfully identify the same potential biomarkers as spin-echo (1)H NMR spectra in which broad lines are suppressed via T2 relaxation editing. Finally, we applied these methods for identification of the metabolic phenotype of patients with type 2 diabetes. This "virtual" relaxation-edited spectroscopy (RESY) approach can be particularly useful for high-throughput screening of complex mixtures such as human plasma and may be useful for extraction of latent biochemical information from legacy or archived NMR data sets for which only standard 1D data sets exist.     

Kinetic and J-resolved statistical total correlation NMR spectroscopy approaches to structural information recovery in complex reacting mixtures: application to acyl glucuronide intramolecular transacylation reactions

We demonstrate here a new variant on a statistical spectroscopic method for recovering structural information on unstable intermediates formed in reaction mixtures. We exemplify this approach with respect to the internal acyl migration reactions of 1-beta-O-acyl glucuronides (AGs), which rearrange at neutral or slightly alkaline pH on a minute to hour time scale to yield a series of positional glucuronide ring isomers and alpha/beta anomers from the 1-beta (starting material), i.e. 2-beta, 2-alpha, 1-alpha, 3-beta, 3-alpha, and 4-beta, 4-alpha isomers together with the aglycon and alpha- and beta-glucuronic acid hydrolysis products. Multiple sequential 800 MHz cryoprobe (1)H NMR spectra (1D and 2D J-resolved, JRES) were collected on a 5.1 mM solution of a synthetic model drug glucuronide, 1-beta-O-acyl (S)-alpha-methyl phenylacetyl glucuronide (MPG) in 0.1 M sodium phosphate buffer in D2O at pD 7.4 over 18 h to monitor the reaction which leads to the formation of the eight positional isomers and hydrolysis products. As the reaction proceeds and new isomers form, the NMR signal intensities vary accordingly allowing the application of a novel kinetic variant on statistical total correlation spectroscopy (K-STOCSY) method to recover the connectivities between proton signals on the same reacting molecule based on their intensity covariance through time. We performed K-STOCSY analysis on both the standard 1D NMR spectra and the skyline projected singlets of the (1)H-(1)H JRES NMR spectra through time, i.e. the K-JRES-STOCSY experimental variant, which increases the effective spectral dispersion and is ideally suited for the analysis of heavily overlapped spin systems. High statistical correlations were observed between mutarotated alpha- and beta-anomers of individual positional isomers, as well as directly acyl migrated products and anticorrelation observed between signals from compounds that were being depleted as others increased, e.g. between the 1-beta and 2-alpha/2-beta isomers. This statistical kinetic approach enabled the recovery of structural connectivity information on all isomers allowing unequivocal resonance assignment, and this approach to spectroscopic information recovery has wider potential uses in the study of reactions that occur on the second-to-minute time scale in conditions where multiple sequential NMR spectra can be collected. JRES-STOCSY is also of potential use as a method for recovering spectroscopic information in highly overlapped NMR signals and spin systems in other types of complex mixture analysis.     

Application of nonselective 1D (1)H-(31)P inverse NMR spectroscopy to the screening of solutions for the presence of organophosphorus compounds related to the chemical weapons convention

1D nonselective (1)H-(31)P HSQMBC, HSQC, and (31)P decoupled HSQC NMR experiments were applied to the screening of original OPCW proficiency test samples for the presence of organophosphorus (OP) compounds related to the Chemical Weapons Convention. The HSQC and HSQMBC spectra are compared to 1D (1)H NMR spectra with WET solvent suppression and (31)P[(1)H] spectra of the same samples. The 1D nonselective HSQC and HSQMBC experiments are shown to be the most sensitive NMR experiments to selectively screen samples for the presence of organophosphorus(OP) compounds. These experiments are at least three times more sensitive than the (31)P[(1)H] NMR experiment and allow the determination of the number of OP compounds present in the sample and their alkyl group bound to the phosphorus atom. Samples spiked at the 5-10 ppm level can be screened within an hour for the presence of OP compounds, whereas for the (31)P[(1)H] experiments, an overnight acquisition is necessary. The sensitivity of the experiments decreases in the order (31)P decoupled HSQC, HSQMBC, and HSQC. For the different alkyl groups, the sensitivity of these experiments decreases in the order methyl approximately isopropyl > ethyl > propyl.     

Statistical heterospectroscopy, an approach to the integrated analysis of NMR and UPLC-MS data sets: application in metabonomic toxicology studies

Statistical heterospectroscopy (SHY) is a new statistical paradigm for the coanalysis of multispectroscopic data sets acquired on multiple samples. This method operates through the analysis of the intrinsic covariance between signal intensities in the same and related molecules measured by different techniques across cohorts of samples. The potential of SHY is illustrated using both 600-MHz 1H NMR and UPLC-TOFMS data obtained from control rat urine samples (n = 54) and from a corresponding hydrazine-treated group (n = 58). We show that direct cross-correlation of spectral parameters, viz. chemical shifts from NMR and m/z data from MS, is readily achievable for a variety of metabolites, which leads to improved efficiency of molecular biomarker identification. In addition to structure, higher level biological information can be obtained on metabolic pathway activity and connectivities by examination of different levels of the NMR to MS correlation and anticorrelation matrixes. The SHY approach is of general applicability to complex mixture analysis, if two or more independent spectroscopic data sets are available for any sample cohort. Biological applications of SHY as demonstrated here show promise as a new systems biology tool for biomarker recovery.     

Metabolic profiling of liver from hypercholesterolemic pigs fed rye or wheat fiber and from normal pigs. High-resolution magic angle spinning 1H NMR spectroscopic study

This study presents the first application of a high-resolution magic angle spinning 1H NMR approach to elucidate the metabolic effects of a hypercholesterolemic condition and two high-fiber diets based on rye and wheat bread, respectively, in intact pig liver biopsy samples. Standard 1D and spin-echo 1H spectra were acquired on a total of 20 biopsy samples, and 2D total correlation spectroscopy experiments were carried out on selected samples for assignment of the observed resonances. Principal component analyses and partial least-squares regression discriminant analysis revealed differences in the hepatic lipid content and choline-containing compounds between normal and hypercholesterolemic pigs. In addition, the results demonstrated that the liver metabolite profile of hypercholesterolemic pigs fed a high-fiber rye bread differed from that of pigs fed high-fiber wheat bread with respect to both the lipoprotein fractions and the choline-containing compounds. These findings suggest that earlier reports on high-fiber rye diet-induced effects on plasma HDL/LDL content partially can be ascribed to effects on liver cholesterol metabolism and that the hepatic phospholipase pathways of phosphatidylcholine breakdown are affected by the high-fiber rye diet.     

Metabolic assessment of human liver transplants from biopsy samples at the donor and recipient stages using high-resolution magic angle spinning 1H NMR spectroscopy

This work presents the first application of high-resolution magic angle spinning (HR-MAS) 1H NMR spectroscopy to human liver biopsy samples, allowing a determination of their metabolic profiles before removal from donors, during cold perfusion, and after implantation into recipients. The assignment of peaks observed in the 1H HR-MAS NMR spectra was aided by the use of two-dimensional J-resolved, TOCSY and 1H-13C HMQC spectra. The spectra were dominated by resonances from triglycerides, phospholipids, and glycogen and from a variety of small molecules including glycerophosphocholine (GPC), glucose, lactate, creatine, acetate, amino acids, and nucleoside-related compounds such as uridine and adenosine. In agreement with histological data obtained on the same biopsies, two of the six livers were found to contain high amounts of triglycerides by NMR spectroscopy, which also indicated that these tissues contained a higher degree of unsaturated lipids and a lower proportion of phospholipids and low molecular weight compounds. Additionally, proton T2 relaxation times indicated two populations of lipids, a higher mobility triglyceride fraction and a lower mobility phospholipid fraction, the proportions of which changed according to the degree of fat content. GPC was found to decrease from the pretransplant to the posttransplant biopsy of all livers except for one with a histologically confirmed high lipid content, and this might represent a biomarker of liver function posttransplantation. NMR signals produced by the liver preservation solution were clearly detected in the cold perfusion stage biopsies of all livers but remained in the posttransplant spectra of only the two livers with a high lipid content and were prominent mainly in the graft that later developed primary graft dysfunction. This study has shown biochemical differences between livers used for transplants that can be related to the degree and type of lipid composition. This technology might therefore provide a novel screening approach for donor organ quality and a means to assess function in the recipient after transplantation.     

Application of a microcoil probe head in NMR analysis of chemicals related to the chemical weapons convention

A 1.7-mm microcoil probe head was tested in the analysis of organophosphorus compounds related to the Chemical Weapons Convention. The microcoil probe head demonstrated a high mass sensitivity in the detection of traces of organophosphorus compounds in samples. Methylphosphonic acid, the common secondary degradation product of sarin, soman, and VX, was detected at level 50 ng (0.52 nmol) from a 30-microL water sample using proton-observed experiments. Direct phosphorus observation of methylphosphonic acid with (31)P{(1)H} NMR experiment was feasible at the 400-ng (4.17 nmol) level. Application of the microcoil probe head in the spiked sample analysis was studied with a test water sample containing 2-10 microg/mL of three organophosphorus compounds. High-quality (1)H NMR, (31)P{(1)H} NMR, 2D (1)H-(31)P fast-HMQC, and 2D TOCSY spectra were obtained in 3 h from the concentrated 1.7-mm NMR sample prepared from 1 mL of the water solution. Furthermore, a 2D (1)H-(13)C fast-HMQC spectrum with sufficient quality was possible to measure in 5 h. The microcoil probe head demonstrated a considerable sensitivity improvement and reduction of measurement times for the NMR spectroscopy in identification of chemicals related to the Chemical Weapons Convention.     

1H MAS NMR (magic-angle spinning nuclear magnetic resonance) techniques for the quantitative determination of hydrogen types in solid catalysts and supports

Distinct hydrogen species are present in important inorganic solids such as zeolites, silicoaluminophosphates (SAPOs), mesoporous materials, amorphous silicas, and aluminas. These H species include hydrogens associated with acidic sites such as Al(OH)Si, non-framework aluminum sites, silanols, and surface functionalities. Direct and quantitative methodology to identify, measure, and monitor these hydrogen species are key to monitoring catalyst activity, optimizing synthesis conditions, tracking post-synthesis structural modifications, and in the preparation of novel catalytic materials. Many workers have developed several techniques to address these issues, including 1H MAS NMR (magic-angle spinning nuclear magnetic resonance). 1H MAS NMR offers many potential advantages over other techniques, but care is needed in recognizing experimental limitations and developing sample handling and NMR methodology to obtain quantitatively reliable data. A simplified approach is described that permits vacuum dehydration of multiple samples simultaneously and directly in the MAS rotor without the need for epoxy, flame sealing, or extensive glovebox use. We have found that careful optimization of important NMR conditions, such as magnetic field homogeneity and magic angle setting are necessary to acquire quantitative, high-resolution spectra that accurately measure the concentrations of the different hydrogen species present. Details of this 1H MAS NMR methodology with representative applications to zeolites, SAPOs, M41S, and silicas as a function of synthesis conditions and post-synthesis treatments (i.e., steaming, thermal dehydroxylation, and functionalization) are presented.     

Application of directly coupled HPLC NMR to separation and characterization of lipoproteins from human serum

Disorders in lipoprotein metabolism are critical in the etiology of several disease states such as coronary heart disease and atherosclerosis. Thus, there is considerable interest in the development of novel methods for the analysis of lipoprotein complexes. We report here a simple chromatographic method for the separation of high-density lipoprotein, low-density lipoprotein, and very low-density lipoprotein from intact serum or plasma. The separation was achieved using a hydroxyapatite column and elution with pH 7.4 phosphate buffer with 100-microL injections of whole plasma. Coelution of HDL with plasma proteins such as albumin occurred, and this clearly limits quantitation of that species by HPLC peak integration. We also show, for the first time, the application of directly coupled HPLC 1H NMR spectroscopy to confirm the identification of the three major lipoproteins. The full chromatographic run time was 90 min with stopped-flow 600-MHz NMR spectra of each lipoprotein being collected using 128 scans, in 7 min. The 1H NMR chemical shifts of lipid signals were identical to conventional NMR spectra of freshly prepared lipoprotein standards, confirming that the lipoproteins were not degraded by the HPLC separation and that their gross supramolecular organization was intact.     

Si cross-polarization magic-angle spinning NMR spectroscopy––an efficient tool for quantification of thaumasite in cement-based materials

²⁹Si{¹H} cross-polarization (CP) magic-angle spinning (MAS) NMR spectroscopy is a powerful and reliable tool for the quantification of thaumasite in cement-based materials. The most efficient method for quantifying thaumasite from ²⁹Si{¹H} CP/MAS NMR spectra is described and it is shown that the method allows detection of thaumasite contents below approximately 10 wt.% with a relative precision of 15% and contents above 10 wt.% with a relative precision of 10%. The applicability of ²⁹Si{¹H} CP/MAS NMR for quantification of thaumasite is demonstrated for different Portland cement pastes and shows that thaumasite contents as low as 0.2–0.5 wt.% can be detected in cementitious systems with low concentrations of paramagnetic impurities. For a Portland cement containing various amounts of limestone dust and stored at 5 °C in a MgSO₄ solution, large quantities of thaumasite have been detected. Furthermore, the quantity of thaumasite is found to be less sensitive to the amount of added limestone dust. For samples of a Portland cement with a fixed content of limestone dust but different quantities of added gypsum, the increased contents of gypsum are observed to result in larger quantities of thaumasite after prolonged hydration.     

Sample classification based on Bayesian spectral decomposition of metabonomic NMR data sets

1H NMR spectra of biofluids provides a wealth of biochemical information on the metabolic status of an organism. Through the application of pattern recognition and classification algorithms, the data have been shown to provide information on disease diagnosis and the beneficial and adverse effects of potential therapeutics. Here, a novel approach is described for identifying subsets of spectral patterns in databases of NMR spectra, and it is shown that the intensities of these spectral patterns can be related to the onset and recovery from a toxic lesion in both a time-related and dose-related fashion. These patterns form a new type of combination biomarker for the biological effect under study. The approach is illustrated with a study of liver toxicity in rats using NMR spectra of urine following administration of a model hepatotoxin hydrazine.     

Concentration-dependent luminescent behaviour of Rhodamine 6G in AlPO4 xerogel monoliths

Homogeneous monolith of AlPO₄ gel doped with Rhodamine 6G (Rh6G) in different dye concentration is prepared by one step process with sol–gel method using the precursors Al(lact)₃ and H₃PO₄. The optical properties of AlPO₄ gel doped with Rh6G have been characterized by UV–vis absorption spectra and fluorescence spectra. Rh6G molecular J-dimers and H-dimers even multimers are analyzed by excitation spectra based on Exciton theory. The structure of AlPO₄ gel doped with Rh6G is investigated by ²⁷Al and ³¹P Magic Angle Solid Nuclear Magnetic Resonance (MAS NMR), ²⁷Al {³¹P} Rotational Echo Double Resonance (REDOR) NMR and ²⁷Al Triple Quantum Magic Angle Spinning (TQ-MAS) NMR. Based on the results of optical spectra and the structural analysis by NMR techniques, The AlPO₄ gel doped with Rh6G dye with molar ratio of Rh6G/Al(lact)₃ of 1 × 10^?4 displays excellent optical properties.     

Structural and spatially resolved studies on the hardening of a commercial resin-modified glass-ionomer cement

A commercial photopolymerizable resin-modified glass-ionomer (Fuji II LC) was studied using a variety of nuclear magnetic resonance (NMR) techniques. ¹H and ¹⁹F stray-field imaging (STRAFI) enabled to follow the acid–base reaction kinetics in self-cured (SC) samples. Gelation and maturation processes with 25 min and 40 h average time constants, respectively, were distinguished. In self- & photo-cured (SPC) samples, two processes were also observed, which occurred with 2 s and 47 s average time constants. ¹H, ²⁷Al and ²⁹Si magic angle spinning (MAS) NMR, ¹³C cross-polarization (CP)/MAS NMR and ²⁷Al multiple quanta (MQ)MAS NMR spectroscopy were used to obtain structural information on the glass and cements that were either SC or SPC. The presence of methacrylate groups was identified in the solid component. Unreacted hydroxyl ethylmethacrylate (HEMA) was detected in self-cured cement. ²⁷Al data showed that approximately 28% and 20% of Al is leached out from glass particles in SC and SPC samples, respectively. The upfield shift detected in ²⁹Si MAS NMR spectra of the cements is consistent with a decrease in the number of Al species in the second coordination sphere of the silicon structures. Scanning electron microscopy (SEM) showed existence of 3D shrinkage of the cement matrix in photo-cured cements.