Nuclear Magnetic Resonance Experiments Research Articles

Fluorine Labeling and 19F NMR Spectroscopy to Study Biological Molecules and Molecular Complexes

High‐resolution nuclear magnetic resonance (NMR) spectroscopy represents a key methodology for studying biomolecules and their interplay with other molecules. Recent developments in labeling strategies have made it possible to incorporate fluorine into proteins and peptides reliably, with manageable efforts and, importantly, in a highly site‐specific manner. Paired with its excellent NMR spectroscopic properties and absence in most biological systems, fluorine has enabled scientists to investigate a rather wide range of scientific objectives, including protein folding, protein dynamics and drug discovery. Furthermore, NMR spectroscopic experiments can be conducted in complex environments, such as cell lysate or directly inside living cells. This review presents selected studies demonstrating how 19F NMR spectroscopic approaches enable to contribute to the understanding of biomolecular processes. Thereby the focus has been set to labeling strategies available and specific NMR experiments performed to answer the underlying scientific objective.

Through-bond and through-space radiofrequency amplification by stimulated emission of radiation

Radio Amplification by Stimulated Emission of Radiation (RASER) is a phenomenon observed during nuclear magnetic resonance (NMR) experiments with strongly negatively polarized systems. This phenomenon may be utilized for the production of very narrow NMR lines, background-free NMR spectroscopy, and excitation-free sensing of chemical transformations. Recently, novel methods of producing RASER by ParaHydrogen-Induced Polarization (PHIP) were introduced. Here, we show that pairwise addition of parahydrogen to various propargylic compounds induces RASER activity of other protons beyond those chemically introduced in the reaction. In high-field PHIP, negative polarization initiating RASER is transferred via intramolecular cross-relaxation. When parahydrogen is added in Earth’s field followed by adiabatic transfer to a high field, RASER activity of other protons is induced via both J-couplings and cross-relaxation. This through-bond and through-space induction of RASER holds potential for the ongoing development and expansion of RASER applications and can potentially enhance spectral resolution in two-dimensional NMR spectroscopy techniques.

A modified surface to volume (SVR) method to calculate nuclear magnetic resonance (NMR) surface relaxivity: Theory and a case study in shale reservoirs

Surface relaxivity (ρ2) is a critical parameter for converting nuclear magnetic resonance (NMR) T2 data to pore size distribution (PSD). The surface-to-volume ratio (SVR) method, known for its simplicity and ease of operation, has been widely used for ρ2 calculation in unconventional reservoirs. However, previous studies often overlooked the equivalence of pore ranges characterized when directly applying the classical SVR model. Moreover, shale reservoirs generally develop layered fractures, whose ρ2 values are different from matrix pores. The logarithmic mean value of the T2 distribution (T2LM) is significantly influenced by layered fractures, therefore, relying solely on the T2LM value of a whole sample under fluid-saturated state will lead to inaccurate ρ2 values of matrix pores, particularly in laminated shales where fractures are well developed. However, insufficient attention has been paid to the effect of fractures on the ρ2 calculation. In this study, a modified SVR method based on the theory of NMR relaxation in partially fluid-saturated pores was proposed to characterize the ρ2 of shale matrix pores. Twenty-four shale core samples from the Shahejie Formation in the Jiyang Depression, China were selected, and subjected to series of NMR experiments at varying oil-bearing conditions, and low-temperature nitrogen adsorption (LTNA) analysis. The results indicate a strong linear correlation (R2 > 0.85) between the inverse T2LM (1/T2LM) and the inverse fluid saturation (1/f) when oil molecules across the entire surface layer participate in the exchange process. For a whole core sample, ρ2 values obtained using the modified SVR model are higher than those obtained using the classical SVR model, especially in samples with numerous fractures. The modified SVR method effectively reduces the impact of fractures on the characterization of ρ2 of matrix pores. For shale pore ρ2 characterization, the classical SVR model may be more suitable for pores smaller than 300 nm, with a recommended T2 range of <33 ms. Additionally, ρ2 values for different pore ranges (<25 nm, 25–100 nm, and >100 nm) within individual samples were estimated. It is found that the ρ2 values of smaller pores is greater than those of larger pores, which may be due to differences in mineralogy of the pores across various size ranges. The small pores are more associated with clay minerals while large pores are surrounded by quartz and rigid minerals. In addition, ρ2 is lower in larger pores and fractures that do not contain organic matter and clays, thus the underestimation of ρ2 by the classical SVR method can be corrected by modified SVR method. This study represents the first attempt to examine ρ2 variations across different pore ranges in shale reservoirs. The methodology presented can be applied to other formations, enhancing NMR data application in both laboratory settings and well logging.

Water-soluble chitosan polymer for enhanced oil recovery in the carbonate reservoir

Numerous surfactants, nanoparticles and polymers have been developed and utilized for enhanced oil recovery applications, irrespective of their environmental impact. Most of these oilfield chemicals were prepared through multi-steps from fossil-based starting materials using hazardous solvents. However, there is a desire to advance eco-friendly and sustainable materials to reduce CO2 emissions and enhance the sustainability of oil production. Herein, we propose the development of chitosan salt, a solely green polymer, prepared in water in the presence of acetic acid (0.1% v/v). The resulting water-soluble polymer demonstrated excellent stability under reservoir conditions and was tested for enhanced oil recovery in the carbonate reservoir. The oil-wet rock wettability was significantly reduced after using chitosan salt solution. Nuclear magnetic resonance (NMR) experiments were used to show the oil displacement at different pores regions using relaxation time (T2 distribution). Imbibition and coreflooding experiments showed that the chitosan salt is able to increase the enhanced oil recovery by extra 16.2% which surpass the recovery obtained from commercial surfactants such as α-olefin sulfonate (AOS) and cetrimonium bromide (CTAB). This study shows that green materials are promising candidates for oil recovery and upstream applications.

Determining Absolute Configuration of Small Molecules by Diffusion NMR Experiments

Enantiomers are ubiquitous in many areas of science, such as pharmaceuticals, agriculture, and food. Nuclear magnetic resonance (NMR) alone is not able to differentiate enantiomers as their spectra are identical. However, these can be distinguished using chiral auxiliaries (such as chiral complexing agents) that form diastereomeric complexes, but absolute identification is still troublesome, usually requiring a chemical reaction with a chiral derivatizing agent. Here, we propose a new method that uses a hybrid mixture of solvating agents in a simple comparison of diffusion NMR experiments, which can discriminate enantiomers in both frequency and diffusion domains, dubbed CHIMERA (CHIral Micelle Enantiomer Resolving Agent). The new method was assessed for twenty‐three small chiral molecules using a combination of BINOL and (‐)‐DMEB, a chiral surfactant, and initial results indicate that absolute configuration can be obtained from a simple experiment.

Determining Absolute Configuration of Small Molecules by Diffusion NMR Experiments.

Enantiomers are ubiquitous in many areas of science, such as pharmaceuticals, agriculture, and food. Nuclear magnetic resonance (NMR) alone is not able to differentiate enantiomers as their spectra are identical. However, these can be distinguished using chiral auxiliaries (such as chiral complexing agents) that form diastereomeric complexes, but absolute identification is still troublesome, usually requiring a chemical reaction with a chiral derivatizing agent. Here, we propose a new method that uses a hybrid mixture of solvating agents in a simple comparison of diffusion NMR experiments, which can discriminate enantiomers in both frequency and diffusion domains, dubbed CHIMERA (CHIral Micelle Enantiomer Resolving Agent). The new method was assessed for twenty-three small chiral molecules using a combination of BINOL and (-)-DMEB, a chiral surfactant, and initial results indicate that absolute configuration can be obtained from a simple experiment.

Geraniol and hydrophobic geraniol:thymol eutectic mixtures: Structure and thermophysical characterization

The properties of essential oils as food preservatives are well-established. These components have also been used as neoteric solvents to prepare eutectic mixtures. Their high capacity to dissolve poorly water-soluble substances makes them particularly interesting in the biotechnology industry. To implement them, it is essential to determine the structural and chemical-physical properties, as well as develop models to predict the behaviour under any operational condition. Herein, pure geraniol and geraniol/thymol mixtures are studied. Nuclear magnetic resonance experiments and molecular dynamics are used to evaluate the fluid structure. In addition, seven thermophysical properties are measured and correlated. The experimental densities, surface tensions, and viscosities were lower than 952 kg m–3, 33 mN m–1, and 36 mPa s, respectively. These values were adequate for the industrial application of these mixtures. PC-SAFT equation of state well predicted the density with an average deviation of 0.13 %, and the isobaric molar heat capacity with a deviation of 6.9 %. Greater compaction was observed at higher thymol molar ratios and dispersive interactions were predominant. Depending on the composition, geraniol was both an acceptor and donor of hydrogen bonds; conversely, thymol acted as a donor. The article provides the necessary knowledge to be able to use these mixtures and design new ones, as ecological solvents in different applications including those in the agri-food sector.

Quelling the Geometry Factor Effect in Quantum Chemical Calculations of 13C NMR Chemical Shifts with the Aid of the pecG-n (n = 1, 2) Basis Sets

A root factor for the accuracy of all quantum chemical calculations of nuclear magnetic resonance (NMR) chemical shifts is the quality of the molecular equilibrium geometry used. In turn, this quality depends largely on the basis set employed at the geometry optimization stage. This parameter represents the main subject of the present study, which is a continuation of our recent work, where new pecG-n (n = 1, 2) basis sets for the geometry optimization were introduced. A goal of this study was to compare the performance of our geometry-oriented pecG-n (n = 1, 2) basis sets against the other basis sets in massive calculations of 13C NMR shielding constants/chemical shifts in terms of their efficacy in reducing geometry factor errors. The testing was carried out with both large-sized biologically active natural products and medium-sized compounds with complicated electronic structures. The former were treated using the computation protocol based on the density functional theory (DFT) and considered in the theoretical benchmarking, while the latter were treated using the computational scheme based on the upper-hierarchy coupled cluster (CC) methods and were used in the practical benchmarking involving the comparison with experimental NMR data. Both the theoretical and practical analyses showed that the pecG-1 and pecG-2 basis sets resulted in substantially reduced geometry factor errors in the calculated 13C NMR chemical shifts/shielding constants compared to their commensurate analogs, with the pecG-2 basis set being the best of all the considered basis sets.

Molecular-Level Understanding of Phase Stability in Phase-Change Nanoemulsions for Thermal Energy Storage by NMR Spectroscopy.

Phase change materials (PCMs) are latent heat storage materials that can store or release thermal energy while undergoing thermodynamic phase transitions. Organic PCMs can be emulsified in water in the presence of surfactants to enhance thermal conductivity and enable applications as heat transfer fluids. However, PCM nanoemulsions often become unstable during thermal cycling. To better understand the molecular origins of phase stability in PCM nanoemulsions, we designed a model PCM nanoemulsion system and studied how the molecular-level environments and dynamics of the surfactants and oil phase changed upon thermal cycling using liquid-state nuclear magnetic resonance (NMR) spectroscopy. The model system used octadecane as the oil phase, stearic acid as the surfactant, and aqueous NaOH as the continuous phase. The liquid fraction of octadecane within the nanoemulsions was quantified noninvasively during thermal cycling by liquid-state 1H single-pulse NMR measurements, revealing the extent of octadecane supercooling as a function of temperature. The mean droplet size of the PCM nanoemulsions, measured by dynamic light scattering (DLS), was correlated with the liquid content of octadecane to explain phase instability in the solid-liquid coexistence region. Quantitative 13C single-pulse NMR experiments established that the carbonyl surfactant head groups were present in multiple distinct environments during thermal cycling. After repeated thermal cycling, the 13C signal intensity of the carbonyl surfactant head groups decreased, indicating that the surfactant head groups lost molecular mobility. The results explain, in part, the origin of phase instability of PCM nanoemulsions upon thermal cycling.

Synthesis and characterization of a chiral spirobifluorene cyclic hexamer

Abstract A new chiral macrocycle 1-[6], consisting of six chiral spirobifluorene units linked in a cyclic arrangement, was successfully synthesized via a homo-coupling of the corresponding acyclic trimer. Notably, this chiral cyclic hexamer exhibited flexibility in solution, and both low-temperature nuclear magnetic resonance (NMR) experiments and density functional theory (DFT) calculations suggest that its stable structure is not the hexagonal D6-symmetric structure but rather the figure-eight-shaped D2-symmetric structure. In this D2-symmetric structure, the opposing bifluorenyl units are π-stacked, and it is suggested that this π-stacked region undergoes rapid dissociation and reformation at room temperature. The HOMO is distributed over the π-stacked region, with a HOMO-LUMO gap of 4.02 eV. The macrocycle exhibited strong violet fluorescence. The absorption dissymmetry factor |gabs| was 7.4 × 10−3, which is larger compared to the series of chiral smaller macrocycles 1-[n] (n = 3 to 5). Additionally, the CPL efficiency, indicated by a BCPL value of 189 M−1 cm−1, is relatively high among chiral organic luminescent molecules.

Integrative Modeling of Protein-Polypeptide Complexes by Bayesian Model Selection using AlphaFold and NMR Chemical Shift Perturbation Data.

Protein-polypeptide interactions, including those involving intrinsically-disordered peptides and intrinsically-disordered regions of protein binding partners, are crucial for many biological functions. However, experimental structure determination of protein-peptide complexes can be challenging. Computational methods, while promising, generally require experimental data for validation and refinement. Here we present CSP_Rank, an integrated modeling approach to determine the structures of protein-peptide complexes. This method combines AlphaFold2 (AF2) enhanced sampling methods with a Bayesian conformational selection process based on experimental Nuclear Magnetic Resonance (NMR) Chemical Shift Perturbation (CSP) data and AF2 confidence metrics. Using a curated dataset of 108 protein-peptide complexes from the Biological Magnetic Resonance Data Bank (BMRB), we observe that while AF2 typically yields models with excellent consistency with experimental CSP data, applying enhanced sampling followed by data-guided conformational selection routinely results in ensembles of structures with improved agreement with NMR observables. For two systems, we cross-validate the CSP-selected models using independently acquired nuclear Overhauser effect (NOE) NMR data and demonstrate how CSP and NMR can be combined using our Bayesian framework for model selection. CSP_Rank is a novel method for integrative modeling of protein-peptide complexes and has broad implications for studies of protein-peptide interactions and aiding in understanding their biological functions.

Potassium- and Lithium-Ammonia Intercalation into Excitonic Insulator Candidate Ta2NiSe5.

Two new reduced phases derived from the topical excitonic insulator candidate Ta2NiSe5 have been synthesized via the intercalation of lithium and potassium from solutions of the metals in liquid ammonia. Li(NH3)Ta2NiSe5 and KTa2NiSe5 both crystallize in orthorhombic space group Pmnb with the following lattice parameters: a = 3.5175(1) Å, b = 18.7828(7) Å, and c = 15.7520(3) Å and a = 3.50247(3) Å, b = 13.4053(4) Å, and c = 15.7396(2) Å, respectively. They have increased unit cell volumes of 48% and 31%, respectively, relative to that of Ta2NiSe5. Significant rearrangement of the transition metal selenide layers is observed in both intercalates compared to the parent phase. In Li(NH3)Ta2NiSe5, neutron diffraction experiments confirm the location of the light Li, N, and H atoms, and solid-state nuclear magnetic resonance (NMR) experiments show that H, N, and Li each occupy a single environment at ambient temperature on the NMR time scale. Magnetometry data show that both intercalates have increased magnetic susceptibilities relative to that of Ta2NiSe5, consistent with the injection of electrons during intercalation and an enhancement of the Pauli paramagnetism.

NMR Spectroscopy and Multiscale Modeling Shed Light on Ion-Solvent Interactions and Ion Pairing in Aqueous NaF Solutions.

The balance between ion solvation and ion pairing in aqueous solutions modulates chemical and physical processes from catalysis to protein folding. Yet, despite more than a century of investigation, experimental determination of the distribution of ion-solvation and ion-pairing states remains elusive, even for archetypal systems like aqueous alkali halides. Here, we combine nuclear magnetic resonance (NMR) spectroscopy and multiscale modeling to disentangle ion-solvent interactions from ion pairing in aqueous sodium fluoride solutions. We have developed a high-accuracy method to collect experimental NMR resonance frequencies for both ions as functions of temperature and concentration. Comparison of these data with resonance frequencies for nonassociating salts allows us to differentiate the influence of solvation and ion pairing on NMR spectra. These high-quality experimental NMR data are used to validate our modeling framework comprising polarizable force field molecular dynamics (MD) simulations and quantum chemical calculations of NMR resonance frequencies. Our experimental and theoretical resonance frequency shifts agree over a wide range of temperatures and concentrations. Structural analysis reveals how both trends are dominated by interactions with water molecules. For the more sensitive 19F nucleus, the NMR resonance frequency decreases as hydrogen bonds between fluoride and water molecules are reduced in number with increased temperature and molality. Through a detailed analysis of the theoretical NMR resonance frequencies for both ions, we show that NMR spectroscopy can distinguish both contact ion pairs and single-solvent-separated ion pairs from free ions. This quantitative framework can be applied directly to other systems.

Polyphosphonate covalent organic frameworks

Herein, we report polyphosphonate covalent organic frameworks (COFs) constructed via P-O-P linkages. The materials are synthesized via a single-step condensation reaction of the charge-assisted hydrogen-bonded organic framework, which is constructed from phenylphosphonic acid and 5,10,15,20‐tetrakis[p‐phenylphosphonic acid]porphyrin and is formed by simply heating its hydrogen-bonded precursor without using chemical reagents. Above 210 °C, it becomes an amorphous microporous polymeric structure due to the oligomerization of P-O-P bonds, which could be shown by constant-time solid-state double-quantum 31P nuclear magnetic resonance experiments. The polyphosphonate COF exhibits good water and water vapor stability during the gas sorption measurements, and electrochemical stability in 0.5 M Na2SO4 electrolyte in water. The reported family of COFs fills a significant gap in the literature by providing stable microporous COFs suitable for use in water and electrolytes. Additionally, we provide a sustainable synthesis route for the COF synthesis. The narrow pores of the COF effectively capture CO2.

Antibacterial and Antioxidant Activities of Hydroalcoholic and Phenolic Extracts from Ternstroemia dentisepala and T. lineata Leaves.

Traditional Mexican medicine commonly uses infusions of Ternstroemia spp. to treat insomnia, injuries, and infections. The antibacterial activities of Ternstroemia dentisepala and Ternstroemia lineata were evaluated for the first time against a panel of Gram-positive and Gram-negative bacteria that have implications for human health, including Enterococcus faecalis, Streptococcus agalactiae, Streptococcus pyogenes, Salmonella typhi, Pseudomonas aeruginosa, and Vibrio parahaemolyticus. Furthermore, the scavenging potential of the hydroalcoholic (HAEs) and total phenolic extracts (TPEs) from the leaves of both plants by a 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assay (ABTS•+) was determined. Also, the total phenolic contents of the HAEs using the Folin-Ciocalteu reagent were assayed. T. dentisepala HAE and TPE were active against all bacterial strains tested, with a minimum inhibitory concentration between 1.0 and 6.0 mg/mL, with the last one being the most active. However, the T. lineata extracts only demonstrated effectiveness against S. typhi and P. aeruginosa. The TPEs from T. dentisepala and T. lineata improved the activity by approximately 30% in all bacteria tested in comparison with the HAEs. The T. dentisepala HAE had a higher total phenolic content than the T. lineata extract, which was consistent with its ABTS•+-scavenging activity. The two HAEs had different chemical profiles, mostly because of the types and amounts of phenolic compounds they contained. These profiles were obtained using thin-layer chromatography (TLC), high-performance liquid chromatography (HPLC), and proton nuclear magnetic resonance (1H NMR) experiments.

Mechanistic Analysis of 5-Hydroxy γ-Pyrones as Michael Acceptor Prodrugs.

Substituted 5-hydroxy γ-pyrones have shown promise as covalent inhibitor leads against cysteine proteases and transcription factors, but their hydrolytic instability has hindered optimization efforts. Previous mechanistic proposals have suggested that these molecules function as Michael acceptor prodrugs, releasing a leaving group to generate an o-quinone methide-like structure. Addition to this electrophile of either water or an active site cysteine was purported to lead to inhibitor hydrolysis or enzyme inhibition, respectively. Through the use of kinetic nuclear magnetic resonance experiments, Hammett analysis, kinetic isotope effect studies, and density functional theory calculations, our findings suggest that enzyme inhibition and hydrolysis proceed by distinct pathways and are differentially influenced by substituent electronics. This mechanistic revision helps enable a more rational optimization for this class of promising compounds.

The Persistence of Hydrogen Bonds in Pyrimidinones: From Solution to Crystal.

Pyrimidinone scaffolds are present in a wide array of molecules with synthetic and pharmacological utility. The inherent properties of these compounds may be attributed to intermolecular interactions analogous to the interactions that molecules tend to establish with active sites. Pyrimidinones and their fused derivatives have garnered significant interest due to their structural features, which resemble nitrogenous bases, the foundational building blocks of DNA and RNA. Similarly, pyrimidinones are predisposed to forming N-H···O hydrogen bonds akin to nitrogenous bases. Given this context, this study explored the supramolecular features and the predisposition to form hydrogen bonds in a series of 18 substituted 4-(trihalomethyl)-2(1H)-pyrimidinones. The formation of hydrogen bonds was observed in solution via nuclear magnetic resonance (NMR) spectroscopy experiments, and subsequently confirmed in the crystalline solid state. Hence, the 18 compounds were crystallized through crystallization assays by slow solvent evaporation, followed by single-crystal X-ray diffraction (SC-XRD). The supramolecular cluster demarcation was employed to evaluate all intermolecular interactions, and all crystalline structures exhibited robust hydrogen bonds, with an average energy of approximately -21.64 kcal mol-1 (∼19% of the total stabilization energy of the supramolecular clusters), irrespective of the substituents at positions 4, 5, or 6 of the pyrimidinone core. To elucidate the nature of these hydrogen bonds, an analysis based on the quantum theory of atoms in molecules (QTAIM) revealed that the predominant intermolecular interactions are N-H···O (average of -16.55 kcal mol-1) and C-H···O (average of -6.48 kcal mol-1). Through proposing crystallization mechanisms based on molecular stabilization energy data and contact areas between molecules and employing the supramolecular cluster and retrocrystallization concepts, it was determined that altering the halogen (F/Cl) at position 4 of the pyrimidinone nucleus modifies the crystallization mechanism pathway. Notably, the hydrogen bonds present in the initial proposed steps were confirmed by 1H NMR experiments using concentration-dependent techniques.

Adsorption and retention of fracturing fluid and its impact on gas transport in tight sandstone with different clay minerals

To elucidate the adsorption characteristics and retention mechanisms of fracturing fluids in diverse clay minerals, we conducted on-line nuclear magnetic resonance (NMR) and atomic force microscopy (AFM) experiments. The depth and extent of solid phase damage are determined by the ratio between the size of fine fractions in fracturing fluid residue and the pore-throat size in experiments. Poor physical properties (K < 0.5 mD) result in a more preferential flow pathway effect during flowback, and the stepwise incremental pressure differential proves to be more effective for the discharge of fracturing fluid in submicron pore throats. The permeability is significantly influenced by the differential distribution of retained fracturing fluid, as supported by direct experimental evidence. The presence of good physical properties (K > 0.5 mD) combined with a scattered distribution of retained fracturing fluid is associated with high gas phase recovery permeability, whereas a continuous sheet-like distribution results in low recovery permeability. The expansive surface area and presence of filamentous illite minerals facilitate the multiple winding and adsorption of fracturing fluids, demonstrating strong hydrogen-bonding, multi-layering and multiple adsorption properties. The geological characteristics of the main gas formations exhibit significant variation, and the severity of damage caused by fracturing fluids occurs in diverse sequences. To address this issue, a differentiated strategy for optimizing fracturing fluids has been proposed.

Displacement-imbibition coupling mechanisms between matrix and complex fracture during injecting-shut in-production process using pore-scale simulation and NMR experiment

Displacement-imbibition coupling oil production, which involves adjusting the operations of a well group (water injection huff and puff in one well, continuous oil production in another well), is an effective technique for enhancing oil recovery (EOR) for tight oil reservoirs. However, the research on displacement-imbibition coupling mechanisms is still missing, especially the combination of pore-scale numerical simulations and physical experiments. In this paper, a novel self-designed displacement-imbibition online nuclear magnetic resonance (NMR) experiment technique is developed. The effect of changes in fracture morphology on the mechanism of displacement-imbibition (oil saturation field and oil displacement efficiency) has been investigated. Based on the fracture morphology of physical experiments, a pore-scale model is established and solved by the finite element method. The pore-scale oil-water two-phase displacement-imbibition during the injecting-shut in-production process is simulated. The results show that the pressure oscillation makes the counter-current imbibition and co-current imbibition occur simultaneously in the matrix pores, which promotes oil recovery in matrix-fracture systems. The strength of displacement and imbibition effects is affected by the complexity of fractures. When the fracture complexity increases, the imbibition of the dense part of the fracture is stronger, while the displacement effect is weakened.

Abietane diterpenoid salvipisone: Structure elucidation, conformational analysis, and antimicrobial activity

Salvipisone, an abietane diterpenoid, was isolated from the roots of Salvia aethiopis L. using a combination of chromatographic separation techniques. Its structure was elucidated employing a variety of spectroscopic methods, including ultraviolet (UV), infrared (IR), 1D and 2D nuclear magnetic resonance (NMR), and mass spectrometry. A comprehensive analysis of experimental 1D and 2D NMR data, incorporating 1H iterative full spin analysis and higher-order multiplet simulation, revealed discrepancies with previously reported NMR data for this compound. Detailed examination of the 1H NMR spectrum's spin-spin coupling structure provided insights into the relative populations of the three staggered conformers around the C12–C13 bond at room temperature. The conformer populations were determined using the relationship between the experimental values for vicinal coupling constants and the estimated ones for the preferred staggered conformers, yielding a ratio of approx. 80:4:16 for anti, gauche, and gauche′-conformers, respectively. Possible reasons for the observed unevenly populated staggered conformers were also discussed. The antimicrobial activity of salvipisone was evaluated using in vitro and in silico methods. In vitro antimicrobial assays demonstrated that salvipisone exhibited the highest inhibitory effect against Staphylococcus aureus ATCC and Enterococcus faecalis ATCC (MIC = 3.125 μg/mL), followed by Bacillus subtilis ATCC (MIC = 6.25 μg/mL). In contrast, concentrations above 100 μg/mL were necessary to inhibit Escherichia coli ATCC and Klebsiella pneumoniae ATCC. In silico studies suggested that salvipisone, particularly its keto form, has significant potential for inhibiting the enzyme gyrase, highlighting its mechanism of antimicrobial action. Our findings highlight the promising antimicrobial and inhibitory properties of salvipisone, particularly against Gram-positive bacteria such as S. aureus.