NMR Detection Research Articles

Continuous Delivery of Hyperpolarized Xenon-129 Gas Using a "Stopped-Flow" Clinical-Scale Cryogen-Free Hyperpolarizer.

In 2022, the FDA approved hyperpolarized (HP) 129Xe gas as an inhalable contrast agent for functional lung imaging. For clinical imaging, HP 129Xe is usually given as a bolus inhalation. However, for preclinical applications (e.g., pulmonary imaging in small rodents), the continuous delivery of HP 129Xe is greatly desired to enable MRI scanning under conditions of physiological continuous animal breathing patterns. Moreover, HP 129Xe gas can be utilized for other applications including materials science and bioanalytical chemistry, where a continuous flow of hyperpolarized gas through an NMR sample over several minutes is also desired for sensing of 129Xe inside an NMR spectrometer. 129Xe is often hyperpolarized using continuous-flow spin-exchange optical pumping, which employs a lean (1-2%) mixture of Xe and a carrier gas (e.g., He and N2). The low Xe concentration in the produced output reduces the NMR detection sensitivity, and thus, Xe cryo-collection is typically employed to achieve near-100% pure gas-phase Xe before administration to the sample or subject. However, the need for cryo-collection undermines a key advantage of continuous-flow production, i.e., the continuous flowing in a hyperpolarizer HP 129Xe gas is trapped inside the hyperpolarizer, and the produced HP 129Xe gas is released at once when the production cycle (30-60 min) is completed. An alternative HP 129Xe production technology employs a "stopped-flow" approach, where a batch of HP gas is hyperpolarized over time and quickly released from a hyperpolarizer. Here, a clinical-scale "stopped-flow" 129Xe hyperpolarizer was employed to hyperpolarize a 1.3 L-atm batch of 50:50 Xe:N2 gas mixture inside a glass cell with an ultralong lifetime of the HP 129Xe state (T1 > 2 h). The produced HP 129Xe gas was slowly delivered into a 5 mm NMR tube via PEEK tubing under a wide range of gas flow rates: 3-180 standard cubic centimeters per minute (sccm). The polarization of the gas ejected from the hyperpolarizer was quantified using in situ low-field NMR polarimetry and additionally verified using a 0.35 T clinical MRI scanner. Continuous-flow delivery of HP 129Xe was demonstrated for up to 15 min with a gas flow rate of 45-150 sccm over a 2.5-m length of PEEK tubing, suffering only small losses in 129Xe polarization. These observations are additionally supported by 129Xe relaxation measurements inside the PEEK tubing employed for gas delivery and the 5 mm NMR tube employed for polarimetry. 129Xe polarization of 16-19% was obtained in the delivered gas, starting with an "in-polarizer" 129Xe polarization of 19%. We envision that this method can be employed for on-demand cryogen-free delivery of hyperpolarized gas using "stopped-flow" 129Xe hyperpolarizers for a broad range of applications, from preclinical imaging to biosensors, and to spectroscopy of materials surfaces.

Solid state NMR spectral editing of histidine, arginine and lysine using Hadamard encoding.

The NMR signals from protein sidechains are rich in information about intra- and inter-molecular interactions, but their detection can be complicated due to spectral overlap as well as conformational and hydrogen exchange. In this work, we demonstrate a protocol for multi-dimensional solid-state NMR spectral editing of signals from basic sidechains based on Hadamard matrix encoding. The Hadamard method acquires multi-dimensional experiments in such a way that both the backbone and under-sampled sidechain signals can be decoded for unambiguous editing in the 15N spectral frequency dimension. All multi-dimensional 15N-edited solid-state NMR experiments can be acquired using this strategy, thereby accelerating the acquisition of spectra spanning broad frequency bandwidth. Application of these methods to the ferritin nanocage, reveals signals from N atoms from His, Arg, Lys and Trp sidechains, as well as their tightly bound, ordered water molecules. The Hadamard approach adds to the arsenal of spectroscopic approaches for protein NMR signal detection.

High H2 Solubility of Perfluorocarbon Solvents and Their Use in Reversible Polarization Transfer from para-Hydrogen.

This research uses perfluorocarbons (PFCs) as effective alternatives to traditional toxic solvents in reversible para-hydrogen-induced polarization (PHIP) for NMR signal enhancement. Hydrogen solubility in PFCs is shown here to be an order of magnitude higher than in typical organic solvents by determination of Henry's constants. We demonstrate how this high H2 solubility enables the PFCs to deliver substantial polarization transfer from para-hydrogen, achieving up to 2400-fold signal gains for 1H NMR detection and 67,000-fold (22% polarization) for 15N NMR detection at 9.4 T in substrates such as pyridine and nicotine. Notably, methylperfluorobutylether outperforms catalytic efficiency in methanol-d4 and dichloromethane-d2 for pyridine at low catalyst loadings. This makes PFCs particularly advantageous for applications demanding high NMR sensitivity. With high polarization efficiency and reduced toxicity, PFCs hold strong potential for expanding hyperpolarized NMR applications across the biomedical and analytical fields.

Exploring Novel Minimal-Catalyst Glucose to CO2 Pathways for Hybrid Enzymatic Biofuel Cell Development

Biological fuel (Biofuel) cells employ biocatalysts to perform redox reactions, converting the chemical energy in a substrate into electrical energy. Glucose is an excellent substrate for biofuel cells due to its bioavailability for implanted devices and because the catalysts involved in its breakdown in vivo, which may be isolated and employed in a fuel cell, are relatively well characterized. Enzymatic glucose fuel cells in literature often catalyze only one to two of twelve possible oxidation steps in the complete oxidation of glucose to CO2 due to issues with multi-enzyme systems, including pH incompatibilities, cross-inhibition between reactions, and competition for electrode surface area. Molecular electrocatalysts such as 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) demonstrate superior substrate scope and electron transfer kinetics in oxidation reactions. We demonstrate the use of 4-amino-TEMPO as an oxidative catalyst to replace four of the six enzymes from a previously established enzymatic glucose to CO2 cascade. The proposed hybrid cascade is supported by NMR and MS detection of intermediate and terminal products.

A Universal Plug-and-Play Approach to In Situ Multinuclear Magnetic Resonance Analysis of Electrochemical Phenomena in Commercial Battery Cells.

Advancing electrochemical energy storage devices relies on versatile analytical tools capable of revealing the molecular mechanisms behind the function and degradation of battery materials in situ. The nuclear magnetic resonance phenomenon plays a pivotal role in fundamental studies of energy materials and devices because of its exceptional sensitivity to local environments and the dynamics of many electrochemically relevant elements. The jelly roll architecture is one of the most energy-dense and, therefore, most popular concepts implemented in pouch, prismatic, and cylindrical Li- and Na-ion cells. Such widely commercialized designs, however, represent a significant obstacle for a range of powerful in situ magnetic resonance-based methodologies due to negligible radio frequency electromagnetic field penetration through conductive metal casings and current collectors. In this work, we introduce an experimental setup that enables direct RF wave transmission through the cell terminals and current collectors, and provides efficient excitation and detection of NMR signals. An RF adapter designed as a plug-and-play device effectively turns the battery cell into an NMR probe that can be tuned to a broad range of Larmor frequencies. Due to its exceptional sensitivity, versatility, and multinuclear capability, the proposed methodology is suitable for fundamental research and industrial high-throughput screening applications. In situ NMR lineshapes provide a direct quantitative description of the chemical environments of electrochemically active elements and enable new metrics for the accurate assessment of state-of-charge (SoC) and state-of-health (SoH). Specifically, in commercial pouch cells based on lithium cobalt oxide, lithium nickel manganese cobalt oxide, and sodium nickel iron manganese oxide chemistries, 7Li and 23Na NMR data unambiguously show electrochemical transformations between intercalated and metallic forms of charge carrier ions, and reveal anisotropic magnetic properties of the electrode coating.

Disentangling the Complexity in Protein Complexes Using Complementary Isotope-Labeling and Multiple-Receiver NMR Spectroscopy.

Intrinsically disordered proteins are abundant in eukaryotic systems, but they remain largely elusive pharmacological targets. NMR spectroscopy proved to be a suitable method to study these proteins and their interaction with one another or with drug candidates. Although NMR can give atomistic information about these interplays, molecular complexity due to severe spectral overlap, limited sample stability, and quantity remain an issue and hamper widespread applications. Here, we propose an approach to simultaneously map protein-protein binding sites onto two interacting partners by employing a complementary isotope-labeling strategy and a multiple receiver NMR detection scheme. With one partner being 15N,2H labeled and the interacting one being 13C,1H-labeled, we exploited proton and carbon detection to obtain clean and easily readable information. The method is illustrated with an application to the 50 kDa ternary protein complex formed between the prominent oncogenic transcription factor complex Myc/MAX and the tumor suppressor BRCA1.

Time-resolved NMR detection of prolyl-hydroxylation in intrinsically disordered region of HIF-1α

Prolyl-hydroxylation is an oxygen-dependent posttranslational modification (PTM) that is known to regulate fibril formation of collagenous proteins and modulate cellular expression of hypoxia-inducible factor (HIF) α subunits. However, our understanding of this important but relatively rare PTM has remained incomplete due to the lack of biophysical methodologies that can directly measure multiple prolyl-hydroxylation events within intrinsically disordered proteins. Here, we describe a real-time 13C-direct detection NMR-based assay for studying the hydroxylation of two evolutionarily conserved prolines (P402 and P564) simultaneously in the intrinsically disordered oxygen-dependent degradation domain of hypoxic-inducible factor 1α by exploiting the "proton-less" nature of prolines. We show unambiguously that P564 is rapidly hydroxylated in a time-resolved manner while P402 hydroxylation lags significantly behind that of P564. The differential hydroxylation rate was negligibly influenced by the binding affinity to prolyl-hydroxylase enzyme, but rather by the surrounding amino acid composition, particularly the conserved tyrosine residue at the +1 position to P564. These findings support the unanticipated notion that the evolutionarily conserved P402 seemingly has a minimal impact in normal oxygen-sensing pathway.

Research on remote reference denoising method based on non-coaxial and non-coplanar tunnel NMR detection

Abstract The limited space within the tunnel constrains the size of the antenna for NMR detection, thereby significantly impacting the signal-to-noise ratio (SNR) of NMR signals. Insufficient SNR data poses substantial challenges to obtaining reliable NMR signals. The paper presents a novel approach to address the challenge of strong background noise in tunnel environments and low SNR data by incorporating the ground multi-channel remote reference denoising method into tunnel NMR advance detection. Specifically designed for narrow tunnels, a multi-channel non-coaxial and non-coplanar remote reference denoising method is proposed. Firstly, the effectiveness of the non-coaxial, non-coplanar remote reference denoising method is verified in the laboratory environment. Secondly, the correlation between the detector antenna and the reference antenna is calculated theoretically to ensure the significant correlation between the detector antenna and the reference antenna. Finally, two processing methods of reference denoising and non-reference denoising are carried out respectively by combining the tunnel detection data. By comparing the inversion results and the engineering construction results, the effectiveness of non-coaxial and non-coplanar remote reference denoising methods in tunnel NMR detection is proved, which provides relevant research support for expanding the application of tunnel NMR detection technology.

Solid state NMR spectral editing of histidine, arginine and lysine using Hadamard encoding.

The NMR signals from protein sidechains are rich in information about intra- and inter-molecular interactions, but their detection can be complicated due to spectral overlap as well as conformational and hydrogen exchange. In this work, we demonstrate a protocol for multi-dimensional solid-state NMR spectral editing of signals from basic sidechains based on Hadamard matrix encoding. The Hadamard method acquires multi-dimensional experiments in such a way that both the backbone and under-sampled sidechain signals can be decoded for unambiguous editing in the 15 N spectral frequency dimension. All multi-dimensional 15 N-edited solid-state NMR experiments can be acquired using this strategy, thereby accelerating the acquisition of spectra spanning broad frequency bandwidth. Application of these methods to the ferritin nanocage, reveals signals from N atoms from His, Arg, Lys and Trp sidechains, as well as their tightly bound, ordered water molecules. The Hadamard approach adds to the arsenal of spectroscopic approaches for protein NMR signal detection.

Screening of ent-copalyl diphosphate synthase and metabolic engineering to achieve de novo biosynthesis of ent-copalol in Saccharomyces cerevisiae

The diterpene ent-copalol is an important precursor to the synthesis of andrographolide and is found only in green chiretta (Andrographis paniculata). De novo biosynthesis of ent-copalol has not been reported, because the catalytic activity of ent-copalyl diphosphate synthase (CPS) is very low in microorganisms. In order to achieve the biosynthesis of ent-copalol, Saccharomyces cerevisiae was selected as the chassis strain, because its endogenous mevalonate pathway and dephosphorylases could provide natural promotion for the synthesis of ent-copalol. The strain capable of synthesizing diterpene geranylgeranyl pyrophosphate was constructed by strengthening the mevalonate pathway genes and weakening the competing pathway. Five full-length ApCPSs were screened by transcriptome sequencing of A. paniculata and ApCPS2 had the best activity and produced ent-CPP exclusively. The peak area of ent-copalol was increased after the ApCPS2 saturation mutation and its configuration was determined by NMR and ESI-MS detection. By appropriately optimizing acetyl-CoA supply and fusion-expressing key enzymes, 35.6 mg/L ent-copalol was generated. In this study, de novo biosynthesis and identification of ent-copalol were achieved and the highest titer ever reported. It provides a platform strain for the further pathway analysis of andrographolide and derivatives and provides a reference for the synthesis of other pharmaceutical intermediates.

Development of a Triphenylmethyl Alkylation Pre-Column Derivatization Method for HPLC Quantitative Analysis of Chemically Unstable Halogenated Compounds.

The primary limitations of the quantitative analysis of thermally labile halogenated compounds by traditional gas chromatography (GC) are the inadequacy of identifying the insufficiently volatile impurity (often with a high boiling point) and the difficulty in obtaining a standard substance with a reliable standardized assay. Taking the 4-(Chloromethyl)-5-methyl-1,3-dioxol-2-one (DMDO-Cl, 1) as an example, we reported a triphenylmethanamino-derivatization method to overcome the challenges of the assay determination of such species. During the quantification of 1, the presence of GC-undetectable polymeric impurity 10 poses a critical challenge in assessing the material quality. Moreover, the standard substance of 1 is not available on the market due to its inherent instability during storage and handling, further complicating the quantitative analysis. In this work, a precolumn HPLC-UV derivatization method based on triphenylmethanamino-alkylation was developed to quantitatively analyze 1. The resulting derivative 2 exhibits excellent crystallinity and superior physical and chemical stability and possesses effective chromophores for UV detection. The conversion from analyte 1 to derivative 2 demonstrates desirable reactivity and purity, facilitating quantitative analysis using the external standard method. The chemical derivatization-chromatographic detection method was optimized and validated, demonstrating its high specificity, good linearity, precision, accuracy, and stability. This method offers a valuable alternative to the general quantitative NMR (qNMR) detection technique, which exhibits reduced specificity in the presence of increased levels of impurities in compound 1.

Optimal sensitivity for 1H detected relayed DNP of organic solids at fast MAS

Dynamic nuclear polarization (DNP) combined with high magnetic fields and fast magic angle spinning (MAS) has opened up a new avenue for the application of exceptionally sensitive 1H NMR detection schemes to study protonated solids. Recently, it has been shown that DNP experiments at fast MAS rates lead to slower spin diffusion and hence reduced DNP enhancements for impregnated materials. However, DNP enhancements alone do not determine the overall sensitivity of a NMR experiment. Here we measure the overall sensitivity of one-dimensional 1H detected relayed DNP experiments as a function of the MAS rate in the 20–60 kHz regime using 0.7 mm diameter rotors at 21.2 T. Although faster MAS rates are detrimental for the DNP enhancement on the target material, due to slower spin diffusion, we find that with increasing spinning rates the gain in sensitivity due to 1H line-narrowing and the folding-in of sideband intensity compensates a large part of the loss of overall hyperpolarization. We find that sensitivity depends on the atomic site in the molecule, and is maximised at between 40 and 50 kHz MAS for the sample of L-histidine.HCl·H2O studied here. There is a 10–20 % difference in sensitivity between the optimum MAS rate and the fastest rate currently accessible (60 kHz).

Polarization losses from the nonadiabatic passage of hyperpolarized solutions through metallic components

From complex-mixture analysis to in vivo molecular imaging, applications of liquid-state nuclear spin hyperpolarization have expanded widely over recent years. In most cases, hyperpolarized solutions are generated ex situ and transported from the polarization instrument to the measurement device. The sample hyperpolarization usually survives this transport, since the changes in magnetic fields that are external to the sample are typically adiabatic (slow) with respect to the internal nuclear spin dynamics. The passage of polarized samples through weakly magnetic components such as stainless steel syringe needles and ferrules is not always adiabatic, which can lead to near-complete destruction of the magnetization. To avoid this effect becoming “folklore” in the field of hyperpolarized NMR, we present a systematic investigation to highlight the problem and investigate possible solutions. Experiments were carried out on: (i) dissolution-DNP-polarized [1-13C]pyruvate with NMR detection at 1.4T, and (ii) 1.5-T-polarized H2O with NMR detection at 2.5μT. We show that the degree of adiabaticity of solutions passing through metal parts is intrinsically unpredictable, likely depending on many factors such as solution flow rate, degree of remanent ferromagnetism in the metal, and nuclear spin species. However, the magnetization destruction effects can be suppressed by application of an external field on the order of 0.1–10mT.

11B NMR of the Morphological Evolution of Traditional Chinese Medicine Borax.

This article applies nuclear magnetic resonance technology to the study of boron-containing traditional Chinese medicine, in order to explore the morphological evolution of boron elements in traditional Chinese medicine. Borax is a traditional Chinese medicine with anti-corrosion, anti-inflammatory, antibacterial, and anticonvulsant effects. It is made by boiling, removing stones, and drying borax minerals like borate salts. This article introduces an 11B nuclear magnetic resonance method for identifying and characterizing boron-containing compounds in TCM. We applied this technology to borax aqueous solutions in different chemical environments and found that with boron mixed in the form of SP2 hybridization in equilateral triangles and SP3 hybridization in equilateral tetrahedra, the pH changes in alkaline environments significantly affected the ratio of the two. At the same time, it was found that in addition to the raw material peak, boron signals of other boron-containing compounds were also detected in 20 commercially available boron-containing TCM preparations. These new boron-containing compounds may be true pharmaceutical active ingredients, and adding them directly to the formula can improve quality and safety. This article describes the detection of 11B NMR in boron-containing traditional Chinese medicine preparations. It is simple, non-destructive, and can provide chemical fingerprint studies for boron-containing traditional Chinese medicine.

Quantitative reaction monitoring using parahydrogen-enhanced benchtop NMR spectroscopy.

The parahydrogen-induced polarisation (PHIP) NMR signal enhancement technique is used to study H2 addition to Vaska's complex (trans-[IrCl(CO)(PPh3)2]) with both standard high-field (9.4 T) NMR and benchtop (1 T) NMR detection. Accurate and repeatable rate constants of (0.84 ± 0.03) dm3 mol-1 s-1 and (0.89 ± 0.03) dm3 mol-1 s-1 were obtained for this model system using standard high-field and benchtop NMR, respectively. The high-field NMR approach is shown to be susceptible to systematic errors associated with interference from non-hyperpolarised signals, which can be overcome through a multiple-quantum filtered acquisition scheme. This challenge is avoided when using benchtop NMR detection because the non-hyperpolarised signals are much weaker due to the lower magnetic field, enabling the use of a simpler and more efficient single RF pulse detection scheme. Method validation against several experimental parameters (NMR relaxation, %pH2 enrichment and temperature) demonstrates the robustness of the benchtop NMR approach but also highlights the need for sample temperature control throughout reaction monitoring. A simple temperature equilibration protocol, coupled with use of an insulated sample holder while manipulating the sample outside the spectrometer, is found to provide sufficient temperature stabilisation to ensure that accurate and repeatable rate constants are obtained. Finally, the benchtop NMR reaction monitoring protocol is applied to the analysis of a complex mixture, where multiple reaction products form simultaneously. H2 addition to a mixture of three Vaska's complex derivatives was monitored, revealing the presence of competitive reaction pathways within the mixture.

Improved on-line benchtop 31P NMR reaction monitoring via Multi-Resonance SHARPER.

On-line reaction monitoring of hydrogenation reactions featuring oxygen-sensitive organometallic complexes is done via a 31P benchtop NMR spectrometer using the Multi-Resonance Sensitive Homogeneous And Resolved PEaks in Real time (MR-SHARPER) sequence. Signal enhancement generated by MR-SHARPER enables monitoring of reactivity on the order of minutes that could not be followed with traditional 31P{1H} NMR detection.

NMR detection of the strained metallacycles in organolithiums: theoretical study.

For the first time through quantum chemistry methods, the effective use of 1JCLi spin-spin coupling constants as descriptors for assessing the formation of strained metallacycles is demonstrated. Both acyclic organolithiums and 3- to 7-membered metallacycles are examined. 80 organolithium compounds, including both monomeric and dimeric species, with ligands containing fluorine, nitrogen, oxygen, and carbon (in the form of carbanions), are tested. In general, the 1JCLi values below 12 Hz for monomeric species and below 6 Hz for dimeric species serve as clear indicators of strained monomeric metallacycle formation (for 6Li nuclei). The primary contributor to the overall 1JCLi value is the Fermi-contact term, which correlates directly with the carbon-lithium interatomic distance and allows to distinguish between dimers and monomers.

Dehydrogenation of Aqueous Formic Acid by Iridium and Ruthenium Complexes with Nitrogen‐rich Diamino‐bis(1H‐1,2,4‐triazole)

AbstractThe development of efficient catalysts for the dehydrogenation of formic acid (FA) through molecular level control and precise regulation remains challenging. Herein, we report the newly developed complexes of iridium and ruthenium with nitrogen‐rich ligand for aqueous FA dehydrogenation. The Cp*Ir complex 1 (Cp*=cyclopentadienyl) and Ru(p‐cym) complex 2 (p‐cym=p‐Cymene) were synthesized and fully characterized. The single crystal structures of these complexes have a typical piano‐stool structure. A turnover frequency of 12468 h−1 was achieved for FA dehydrogenation at 90 °C by using complex 1 bearing 3,3’‐diamino‐5,5’‐bis(1H‐1,2,4‐triazole) (DABT). The effects of pH, temperature, concentration of formic acid and amount of catalyst on the reaction kinetics were studied in detail. Kinetic isotope effect (KIE) experiments and 1H NMR detection of reaction revealed that a plausible mechanism involves the decarboxylation of formate and the formation of H2. More importantly, the influence of H2O and H+ was involved in the rate‐determining step. This study demonstrated the effectiveness of nitrogen‐rich ligand for FA dehydrogenation, which would contribute to the development of new and effective hydrogen release systems.

Solid-state 35/37 Cl NMR detection of chlorine atoms directly bound to paramagnetic cobalt(II) ions in powder samples.

We report high-quality solid-state 35/37 Cl NMR spectra for chlorine atoms directly bonded to paramagnetic cobalt(II) ions (high spin S = 3/2) in powered samples of CoCl2 , CoCl2 ·2H2 O, CoCl2 ·6H2 O, and CoCl2 (terpy) (terpy = 2,2':6',2″-terpyridine). Because solid-state 35/37 Cl NMR spectra for paramagnetic cobalt(II) compounds often cover an extremely wide spectral range, they were recorded in this work in the form of variable-offset cumulative spectra. Solid-state 35/37 Cl NMR measurements were performed at three magnetic fields (11.7, 14.1, and 16.5T) and analysis of data yielded information about 35/37 Cl quadrupole coupling and hyperfine coupling tensors in these paramagnetic cobalt(II) compounds. Experimental 35/37 Cl NMR tensors were found to be in reasonable agreement with quantum chemical calculations based on a periodic DFT method implemented in BAND.

An effective method for the direct crystallization of xylonic acid from fermentation broth of agricultural residue hydrolysate

Xylose is the second most abundant sugar in nature and conversion of xylose to xylonic acid (XA) has become a research hotspot in recent years. Xylonic acid can be applied in the equivalent market niche of gluconic acid, due to their similar physical properties. XA bioproduction and application presents certainly a promising approach for xylose valorization in lignocellulose biorefinery, while the XA crystallization is so far still an unattainable goal. In this paper, an original method was proposed and experimentally investigated for the first time to get XA crystals from fermentation broth. In the XA crystallization, methanol was introduced as buffer-cum-solvent. The H2SO4 was dropwise added into the methanol solution dissolving potassium xylonate that is beforehand produced by fermentation. During H2SO4 blending with methanol, the strong and polar H2SO4 leads to XA diffusion and aggregation. Overall, 2.21 g of xylonic acid crystals was obtained per gram of 98% H2SO4 acidification. NMR detection showed that over 99% purity of XA crystals was finally obtained at the yields of 67.2% from xylonate. The integrated process could provide a practicable and effective technology for the high-qualify XA preparation from xylose derivations and lignocellulose biomass biorefinery.