Study on tissue distribution, metabolite profiling, and excretion of [14C]-labeled flonoltinib maleate in rats.

Flonoltinib Maleate (FM) is a dual-target inhibitor that selectively suppresses Janus kinase 2/FMS-like tyrosine kinase 3 (JAK2/FLT3), which is currently in phase I/IIa clinical trial in China for the treatment of myeloproliferative neoplasms (MPNs). In this research, we used [14C]-labeled FM (14C-FM) to investigate the distribution, metabolism, and excretion of FM in rats using High-Performance Liquid Chromatography coupled with High-Resolution Mass Spectrometry/Radioactivity Monitoring (HPLC-HRMS/RAM) and liquid scintillation counter. The results revealed that FM displayed widespread distribution in rats. Furthermore, FM demonstrated rapid clearance without any observed risk of organ toxicity attributed to accumulation. Profiling of FM metabolites in rat plasma, feces, urine, and bile identified a total of 17 distinct metabolites, comprising 7 phase I metabolites and 10 phase II metabolites. The major metabolic reactions involved oxygenation, dealkylation, methylation, sulfation, glucuronidation and glutathione conjugation. Based on these findings, a putative metabolic pathway of FM in rats was proposed. The overall recovery rate in the excretion experiment ranged from 93.04 % to 94.74 %. The results indicated that FM undergoes extensive hepatic metabolism in SD rats, with the majority being excreted through bile as metabolites and ultimately eliminated via feces. A minor fraction of FM (<10 %) was excreted through renal excretion in the form of urine. Integration of the current results with previous pharmacokinetic investigations of FM in rats and dogs enables a comprehensive elucidation of the in vivo ADME processes and characteristics of FM, thereby establishing a solid foundation for subsequent clinical investigations of FM.

Composition of raft-like cell membrane microdomains resistant to styrene-maleic acid copolymer (SMA) solubilization.

An advantageous alternative to the use of detergents in biochemical studies on membrane proteins are the recently developed styrene-maleic acid (SMA) amphipathic copolymers. In our recent study [1] we demonstrated that using this approach, most T cell membrane proteins were fully solubilized (presumably in small nanodiscs), while two types of raft proteins, GPI-anchored proteins and Src family kinases, were mostly present in much larger (>250 nm) membrane fragments markedly enriched in typical raft lipids, cholesterol and lipids containing saturated fatty acid residues. In the present study we demonstrate that disintegration of membranes of several other cell types by means of SMA copolymer follows a similar pattern and we provide a detailed proteomic and lipidomic characterization of these SMA-resistant membrane fragments (SRMs).

Immunochemical characterisation of styrene maleic acid lipid particles prepared from Mycobacterium tuberculosis plasma membrane.

Membrane proteins of Mycobacterium tuberculosis (Mtb) can be targeted for the development of therapeutic and prophylactic interventions against tuberculosis. We have utilized the unique membrane-solubilising properties of the styrene maleic acid copolymer (SMA) to prepare and characterise 'styrene maleic acid lipid particles' from the native membrane of Mtb (MtM-SMALPs). When resolved by SDS-PAGE and visualised with coomassie blue, the molecular weights of Mtb membrane (MtM) proteins solubilised by SMA were mostly in the range of 40-70 kDa. When visualised by transmission electron microscopy, MtM-SMALPs appeared as nanoparticles of discrete shapes and sizes. The discoid nanoparticles exhibited a range of diameters of ~10-90 nm, with largest portion (~61%) ranging from 20-40 nm. MtM proteins of a molecular weight-range overlapping with that of MtM-SMALPs were also amenable to chemical cross-linking, revealing protein complex formation. Characterisation using monoclonal antibodies against seven MtM-associated antigens confirmed the incorporation of the inner membrane protein PRA, membrane-associated proteins PstS1, LpqH and Ag85, and the lipoglycan LAM into MtM-SMALPs. Conversely, the peripheral membrane proteins Acr and PspA were nearly completely excluded. Furthermore, although MtM showed an abundance of Con A-binding glycoproteins, MtM-SMALPs appeared devoid of these species. Immune responses of healthcare workers harbouring 'latent TB infection' provided additional insights. While MtM-SMALPs and MtM induced comparable levels of the cytokine IFN-γ, only MtM-SMALPs could induce the production of TNF-α. Antibodies present in the donor sera showed significantly higher binding to MtM than to MtM-SMALPs. These results have implications for the development of MtM-based immunoprophylaxis against tuberculosis.

Design and synthesis of malonohydrazide based colorimetric receptors for discrimination of maleate over fumarate and detection of F-, AcO- and AsO2- ions.

In this study, we have designed and synthesized two new organic receptors R1 and R2 based on malonohydrazide for the recognition of biologically important anions. The receptor R1 capable of colorimetric discrimination of maleate over fumarate ion, exhibit significant color change from pale yellow to wine red due to intermolecular hydrogen bond between the R1 and maleate ion, supported by 1HNMR titration, where the peak at δ12.0 ppm attributed to the NH proton experiences a downfield shift upon binding with maleate ion. Receptor R1, equipped with two electron-withdrawing NO2 moieties as the chromogenic signaling unit enhance the hydrogen bonding tendency and acidity, and thus when comparing with receptor R2, R1 displayed substantial higher redshift (∆λmax) of 148 nm, 143 nm, and 140 nm towards F-, AcO-, and maleate anion in the DMSO. In addition, the synthesized receptors R1 and R2 are able to detect F-, AcO-, and AsO2- ions as their sodium salts in an aqueous solution with visual color change. Receptor R1 exhibit electrochemical response towards F- and AcO- ions. The receptors R1 and R2 are successfully applied for quantitative detection of F- ion in the toothpaste solution in an aqueous medium. Additionally, R1 and R2 exhibit fluorescence enhancement towards F- and AcO- ions in the DMSO. As well, R1 and R2 demonstrate to be potentially useful colorimetric chemosensor for sensing maleate ion using the test strip. The theoretical calculation based on TD-DFT corroborates well with the experimental results of the receptors R1 and R2 with fluoride, acetate and maleate.

A Transcription Factor-Based Biosensor for Detection of Itaconic Acid.

Itaconic acid is an important platform chemical that can easily be incorporated into polymers and has the potential to replace petrochemical-based acrylic or methacrylic acid. A number of microorganisms have been developed for the biosynthesis of itaconate including Aspergillus terreus, Escherichia coli, and Saccharomyces cerevisiae. However, the number of strains and conditions that can be tested for increased itaconate titers are currently limited because of the lack of high-throughput screening methods. Here we identified itaconate-inducible promoters and their corresponding LysR-type transcriptional regulators from Yersinia pseudotuberculosis and Pseudomonas aeruginosa. We show that the YpItcR/P ccl inducible system is highly inducible by itaconic acid in the model gammaproteobacterium E. coli and the betaproteobacterium Cupriavidus necator (215- and 105-fold, respectively). The kinetics and dynamics of the YpItcR/P ccl inducible system are investigated, and we demonstrate, that in addition to itaconate, the genetically encoded biosensor is capable of detecting mesaconate, cis-, and trans-aconitate in a dose-dependent manner. Moreover, the fluorescence-based biosensor is applied in E. coli to identify the optimum expression level of cadA, the product of which catalyzes the conversion of cis-aconitate into itaconate. The fluorescence output is shown to correlate well with itaconate concentrations quantified using high-performance liquid chromatography coupled with ultraviolet spectroscopy. This work highlights the potential of the YpItcR/P ccl inducible system to be applied as a biosensor for high-throughput microbial strain development to facilitate improved itaconate biosynthesis.

Detection of unexpected frauds: Screening and quantification of maleic acid in cassava starch by Fourier transform near-infrared spectroscopy.

Fourier transform near-infrared (FT-NIR) spectroscopy and chemometrics were adopted for the rapid analysis of a toxic additive, maleic acid (MA), which has emerged as a new extraneous adulterant in cassava starch (CS). After developing an untargeted screening method for MA detection in CS using one-class partial least squares (OCPLS), multivariate calibration models were subsequently developed using least squares support vector machine (LS-SVM) to quantitatively analyze MA. As a result, the OCPLS model using the second-order derivative (D2) spectra detected 0.6%(w/w) adulterated MA in CS, with a sensitivity of 0.954 and specificity of 0.956. The root mean squared error of prediction (RMSEP) was 0.192(w/w, %) by using the standard normal variate (SNV) transformation LS-SVM. In conclusion, the potential of FT-NIR spectroscopy and chemometrics was demonstrated for application in rapid screening and quantitative analysis of MA in CS, which also implies that they have other promising applications for untargeted analysis.

Quantitative Nuclear Magnetic Resonance Spectroscopy Based on PULCON Methodology: Application to Quantification of Invaluable Marine Toxin, Okadaic Acid.

ERETIC2 (Electronic Reference To access In vivo Concentrations 2) based on PULCON (Pulse Length-based Concentration determination) methodology is a quantitative NMR (qNMR) using an external standard. The performance of the PULCON method was assessed using maleic acid (MA). Quantification of the diarrhetic shellfish toxin and okadaic acid by PULCON was successfully consistent with that obtained by a conventional internal standard method, demonstrating that the PULCON method is useful for the quantification of invaluable marine toxins without any contaminations by an internal standard.

Colorimetric and fluorometric discrimination of geometrical isomers (maleic acid vs fumaric acid) with real-time detection of maleic acid in solution and food additives.

Heterobis imine Schiff base probe L is able to discriminate geometrical isomers (maleic acid vs fumaric acid) through sharp colorimetric as well as fluorogenic responses even conspicuous with the naked eye. Colorimetric as well as fluorogenic sensing of maleic acid among various carboxylic acids was also demonstrated in ethanol-buffer medium. Sensing behavior of L was corroborated by (1)H NMR spectra, mass spectrometry, and theoretical calculations. Subsequently sensing behavior of L was used to probe maleic acid in starch rich food samples.

Application of quantitative NMR for purity determination of standard ACE inhibitors.

This study investigated the accuracy of the quantitative NMR method for purity determination of ACE inhibitors reference standards and the discovery of two pairs of new diastereoisomers. Six types of ACE inhibitors, imidapril hydrochloride, benazepril hydrochloride, lisinopril, enalapril maleate, quinapril hydrochloride, and captopril were quantificated and validated for the qNMR method by discussing factors that affect parameters of the qNMR experiment, internal standards, integration, pH-effect, and uncertainty. The results were compared with data obtained by the mass balance method. The study found that maleic acid influenced the quantification of captopril in deuteroxide because of a chemical reaction. The mixtures of the reaction products were isolated by HPLC and structurally elucidated by NMR as two pairs of new diastereoisomers, 1-[(2S,4R)-thio-2-methylpropionyl-5-d-ethanedicarboxylicacid]-L-proline and 1-[(2S,4S)-thio-2-methylpropionyl-5-d-ethanedicarboxylicacid]-L-proline. The results showed that the accuracy and precision of quantitative (1)H NMR spectroscopy satisfied the requirements for quantitative analysis of chemical reference standards and provided a simple, rapid, and reliable method for purity determination of ACE inhibitors systematically.

Adsorption and transport of polymaleic acid on Callovo-Oxfordian clay stone: batch and transport experiments.

Dissolved Organic Matter (DOM) can affect the mobility of radionuclides in pore water of clay-rich geological formations, such as those intended to be used for nuclear waste disposal. The present work studies the adsorption and transport properties of a polycarboxylic acid, polymaleic acid (PMA, Mw=1.9kDa), on Callovo-Oxfordian argillite samples (COx). Even though this molecule is rather different from the natural organic matter found in clay rock, the study of its retention properties on both dispersed and intact samples allows assessing to which extent organic acids may undergo sorption under natural conditions (pH7) and what could be the impact on their mobility. PMA sorption and desorption were investigated in dispersed systems. The degree of sorption was measured after 1, 8 and 21days and for a range of PMA initial concentrations from 4.5×10(-7) to 1.4×10(-3)mol.L(-1). The reversibility of the sorption process was estimated by desorption experiments performed after the sorption experiments. At the sorption steady state, the sorption was described by a two-site Langmuir model. A total sorption capacity of COx for PMA was found to be 1.01×10(-2) mol.kg(-1) distributed on two sorption sites, one weak and one strong. The desorption of PMA was incomplete, independently of the duration of the sorption phase. The amount of desorbable PMA even appeared to decrease for sorption phases from 1 to 21days. To describe the apparent desorption hysteresis, two conceptual models were applied. The two-box diffusion model accounted for intraparticle diffusion and more generally for nonequilibrium processes. The two-box first-order non-reversible model accounted for a first-order non-reversible sorption and more generally for kinetically-controlled irreversible sorption processes. The use of the two models revealed that desorption hysteresis was not the result of nonequilibrium processes but was due to irreversible sorption. Irreversible sorption on the strong site was completed after 1day and represented 96% of the total sorption on this site. On the weak site the irreversible uptake was slower and completed only after 16days but it also dominated the sorption. 85% of the PMA sorbed on the weak site was not desorbable after 21days of sorption. The migration of PMA was studied by applying a hydraulic gradient to a clay core inserted in a stainless steel cell. Breakthrough of polymaleic acid, simulated with a 1D transport model including the two-box first-order non-reversible model, revealed that the mobility of PMA was limited by the same set of reversible/irreversible interactions as observed in the dispersed system. However, to describe efficiently the transport, the total sorption capacity had to be reduced to 33% of the capacity estimated in batch experiments. The irreversible sorption on the weak site was also slower in the intact sample than in the crushed sample. Geometrical constraints would therefore affect both the accessibility to the sorption sites and the kinetics of the irreversible sorption process.

The first insight into the metabolite profiling of grapes from three Vitis vinifera L. cultivars of two controlled appellation (DOC) regions.

The characterization of the metabolites accumulated in the grapes of specific cultivars grown in different climates is of particular importance for viticulturists and enologists. In the present study, the metabolite profiling of grapes from the cultivars, Alvarinho, Arinto and Padeiro de Basto, of two Portuguese Controlled Denomination of Origin (DOC) regions (Vinho Verde and Lisboa) was investigated by gas chromatography-coupled time-of-flight mass spectrometry (GC-TOF-MS) and an amino acid analyzer. Primary metabolites, including sugars, organic acids and amino acids, and some secondary metabolites were identified. Tartaric and malic acids and free amino acids accumulated more in grapes from vines of the DOC region of Vinho Verde than DOC Lisboa, but a principal component analysis (PCA) plot showed that besides the DOC region, the grape cultivar also accounted for the variance in the relative abundance of metabolites. Grapes from the cultivar, Alvarinho, were particularly rich in malic acid and tartaric acids in both DOC regions, but sucrose accumulated more in the DOC region of Vinho Verde.

Identification of putative steroid receptor antagonists in bottled water: combining bioassays and high-resolution mass spectrometry.

Endocrine disrupting chemicals (EDCs) are man-made compounds interfering with hormone signaling and thereby adversely affecting human health. Recent reports provide evidence for the presence of EDCs in commercially available bottled water, including steroid receptor agonists and antagonists. However, since these findings are based on biological data the causative chemicals remain unidentified and, therefore, inaccessible for toxicological evaluation. Thus, the aim of this study is to assess the antiestrogenic and antiandrogenic activity of bottled water and to identify the causative steroid receptor antagonists. We evaluated the antiestrogenic and antiandrogenic activity of 18 bottled water products in reporter gene assays for human estrogen receptor alpha and androgen receptor. Using nontarget high-resolution mass spectrometry (LTQ-Orbitrap Velos), we acquired corresponding analytical data. We combined the biological and chemical information to determine the exact mass of the tentative steroid receptor antagonist. Further MS(n) experiments elucidated the molecule's structure and enabled its identification. We detected significant antiestrogenicity in 13 of 18 products. 16 samples were antiandrogenic inhibiting the androgen receptor by up to 90%. Nontarget chemical analysis revealed that out of 24520 candidates present in bottled water one was consistently correlated with the antagonistic activity. By combining experimental and in silico MS(n) data we identified this compound as di(2-ethylhexyl) fumarate (DEHF). We confirmed the identity and biological activity of DEHF and additional isomers of dioctyl fumarate and maleate using authentic standards. Since DEHF is antiestrogenic but not antiandrogenic we conclude that additional, yet unidentified EDCs must contribute to the antagonistic effect of bottled water. Applying a novel approach to combine biological and chemical analysis this is the first study to identify so far unknown EDCs in bottled water. Notably, dioctyl fumarates and maleates have been overlooked by science and regulation to date. This illustrates the need to identify novel toxicologically relevant compounds to establish a more holistic picture of the human exposome.

[Determination of the total amount of maleic acid and maleic anhydride in starch and its products by high performance liquid chromatography-tandem mass spectrometry].

A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/ MS) method was established for the determination of the total amount of maleic acid and maleic anhydride in starch and its products. The samples were extracted with 50% (v/v) methanol and hydrolyzed by alkaline, then analyzed by HPLC-MS/MS, and quantified with the external standard method. The mass spectrometry was operated with electrospray in negative ionization mode. The multiple reaction monitoring (MRM) mode was employed involving the transition of the precursor ion to two selected product ions, in which one pair was for quantification (m/z 115.0 > 71.1) and the other pair was for identification (m/z 115.0 > 27.1). The results indicated that no significant matrix effect was found for the spiked samples. The recoveries of maleic acid spiked in starch and its products were 80.2%-115.3% at spiked levels of 0.5-1,000 mg/kg. The relative standard deviations (RSDs) were less than 12% (n = 6). The limits of detection (LOD) and quantification (LOQ) were 0.1 and 0.5 mg/kg for maleic acid and maleic anhydride, respectively. The method is rapid, sensitive and reproducible for the determination of the total amount of maleic acid and maleic anhydride in starch and its products and shows great potential for routine analysis

Method validation and determinations of levofloxacin, metronidazole and sulfamethoxazole in an aqueous pharmaceutical, urine and blood plasma samples using quantitative nuclear magnetic resonance spectrometry.

Selective, rapid and accurate quantitative proton nuclear magnetic resonance (qHNMR) method for the determination of levofloxacin, metronidazole benzoate and sulfamethoxazole in aqueous solutions was developed and validated. The method was successfully applied to the determinations of the drugs and their admixtures in pharmaceutical, urine and plasma samples. Maleic acid and sodium malate were used as internal standards. Effect of temperature on spectral measurements was evaluated. Linear dynamic ranges of 0.50-68.00, 0.13-11.30 and 0.24-21.00 mg per 0.60 mL solution were obtained for levofloxacin, metronidazole benzoate and sulfamethoxazole, respectively. Average recovery % in the range of 96.00-104.20 ± (0.17-2.91) was obtained for drugs in pure, pharmaceutical, plasma and urine samples. Inter and intra-day analyses gave average recoveries % in the ranges 96.10-98.40 ± (1.68-2.81) and 96.00-104.20 ± (0.17-2.91), respectively. Instrumental detection limits ≤0.03 mg per 0.6 mL were obtained for the three drugs. Developed method has demonstrated high performance characteristics for analyzing investigated drugs and their admixtures. Student t-test at 95% confidence level revealed insignificant bias between the real and measured contents of investigated drugs in pure, pharmaceutical, urine and plasma samples and its admixtures. Application of the statistical F-test revealed insignificant differences in precisions between the developed method and arbitrary selected reference methods.

(rac)-1,1'-binaphthyl-based simple receptors designed for fluorometric discrimination of maleic and fumaric acids.

(rac)-1,1'-Binaphthyl-based simple receptors 1 and 2 have been designed, synthesized and studied theoretically. The receptors utilize naphthyridine as the binding motifs for complexation of dicarboxylic acids in CHCl(3). The emission of the BINOL moiety was monitored experimentally to ascertain the selectivity and sensitivity of the receptors. Receptor 1 distinguishes maleic acid from isomeric fumaric acid by exhibiting different fluorescence behavior and demonstrates stronger binding in the excited state. Modulation of the binding sites of 1 leads to a new receptor structure 2, which was found to be less efficient in distinguishing maleic from fumaric acid, fluorometrically. Both 1 and 2 also recognize other hydroxy di- and tricarboxylic acids. The binding interactions were monitored by (1)H NMR, fluorescence and UV-vis spectroscopic methods. Structures of apo-hosts, guests and host-guest complexes were determined using force-field based conformational searching. Low energy ensembles were grouped into geometrically similar families, and low energy structures from each family were verified using B3LYP/6-31G*/PB-SCRF(CHCl(3)) calculations. The atomistic calculations provide insight into the differential dicarboxylic acid binding behavior of receptors 1 and 2.

Determination of fumaric and maleic acids with stacking analytes by transient moving chemical reaction boundary method in capillary electrophoresis.

The paper presents an on-line transient moving chemical reaction boundary (MCRB) method for simply but efficiently stacking analytes in capillary electrophoresis (CE). The CE technique was developed for a rapid determination of fumaric and maleic acid. Based on the theory of MCRB, Effects of several important factors such as the pH and concentration of running buffer and the conditions of stacking analytes were investigated to acquire the optimum conditions. The optimized separations were carried out in a 20 mmol/L sulphate neutralized with ethylenediamine to pH 6.0 electrolytes using a capillary coated with poly (diallyldimethylammonium chloride) and direct UV detection at 214 nm. The optimized preconcentrations were carried out in 50 mmol/L borax (pH 9.0). The calibration curves were linear in the concentration range of 1.0×10⁻7-1.0×10⁻4 mol/L and 5.0×10⁻7-1.0×10⁻4 mol/L for fumaric and maleic acid with correlation coefficients higher than 0.9991. The detection limits were 5.34×10⁻8 mol/L for fumaric acid and 1.92×10⁻7 mol/L for maleic acid. This method was applied for determination of fumaric acid in apple juice and of fumaric and maleic acid in dl-malic, the recovery tests established for real samples were within the range 95-105%. This work provided a valid and simple approach to detect fumaric and maleic acid.

Characterization of composites based on expanded polystyrene wastes and wood flour.

This paper aims to evaluate the potential for the use of recycled expanded polystyrene and wood flour as materials for the development of wood plastic composites. The effects of wood flour loading and coupling agent addition on the mechanical properties and morphology of wood thermoplastic composites were examined. In addition, a methodology for the thermo-mechanical recycling of expanded polystyrene waste was developed. The results show that the mechanical properties decreased as the wood flour loading increased. On the other hand, the use of poly(styrene-co-maleic anhydride), SMA, as a coupling agent improved the compatibility between the wood flour and polystyrene matrix and the mechanical properties subsequently improved. A morphological study revealed the positive effect of the coupling agent on the interfacial bonding. The density values obtained for the composites were compared with the theoretical values and showed agreement with the rule of mixtures. Based on the findings of this work, it appears that both recycled materials can be used to manufacture composites with high mechanical properties and low density.

An HPLC with evaporative light scattering detection method for the quantification of PEGs and Gantrez in PEGylated nanoparticles.

A rapid and precise HPLC method with evaporative light scattering detection (ELSD) for the separation and quantification of polyethyleneglycol 2000 (PEG 2000), polyethyleneglycol 6000 (PEG 6000) and poly(methyl vinyl ether-co-maleic anhydride) (Gantrez) in a nanosized pharmaceutical formulation has been developed. Separation was carried out on a PL aquagel-OH 30,8 microm column (300 mm x 7.5 mm), in a gradient elution with methanol-water as mobile phase at a flow rate of 1 ml/min. Quantification was determined in supernatants of PEGylated nanoparticles and the quantification limits were found to be 0.075 mg/ml for polyethyleneglycols and 0.25 mg/ml for Gantrez. The precision did not exceed 8% and accuracy range for PEGs (-11.50 and 10.61%) and Gantrez (-12.18 and 14.81%) were always within the acceptable limits. The amount of polyethyleneglycol associated to nanoparticles was also calculated by a Nuclear Magnetic Resonance Method ((1)H NMR). Likely, for both PEGs, a good relationship between both techniques was found. In summary, the developed HPLC technique provides an alternative for the routine and rapid analysis of PEGs and Gantrez in nanoparticle formulations.

Derivatisation of carboxylic acid groups in pharmaceuticals for enhanced detection using liquid chromatography with electrospray ionisation tandem mass spectrometry.

Tris(2,4,6-trimethoxyphenyl)phosphonium propylamine bromide (TMPP) has been used for the derivatisation of maleic, fumaric, sorbic and salicylic acids to facilitate determination using liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/ESI-MS/MS) in positive ion mode. Detection limits, achieved using multiple reaction monitoring mode, were 2, 4, 0.4 and 540 fmol (5 muL injection) for derivatised fumaric, sorbic, maleic and salicylic acids, respectively. In comparison, detection limits achieved in negative ion mode for the underivatised acids were 24, 51, 2, and 117 fmol, respectively. The method was successfully used for the determination of sorbic acid in a sample of Panadol. The derivatisation of salicylic acid was not as successful, probably due to poor reaction efficiency.

Double-quantum filtered 1H MAS NMR spectra.

It is shown that straightforward double-quantum filtered (1)H MAS NMR experiments yield spectral lineshapes that permit to estimate the minimum number of (1)H spins in a cluster. The approach may offer an alternative to multiple-quantum experiments for the characterisation of (1)H spin clusters of moderate size. The duration of the double-quantum excitation period has to be chosen suitably, it is necessary to find a practical compromise between optimum double-quantum filtration efficiency and optimum information content of the spectral lineshapes. Some (1)H MAS NMR experiments on partially deuterated maleic acid are reported as well as numerical simulations.