Chromatography Experiments Research Articles

Decolorization of anthraquinone dye by Brevibacillus laterosporus R81 via polysaccharides excreted

AbstractThe aim of this research was to assess the efficacy of different microbial strains in the decolorization of anthraquinone dyes. Strain R81 was obtained from a textile company's wastewater discharge for its remarkable ability to decolorize reactive blue 19 (RB19). By employing physiological and biochemical analyses, along with 16S rRNA gene sequencing, strain R81 was determined to be Brevibacillus laterosporus. After optimization, the decolorization rate achieved a peak of 86.24% over a 48‐h timeframe, utilizing an initial dye concentration of 100 mg L–1. The decolorization capacity of strain R81 was observed to be impeded by heightened levels of salt and temperature in culture solutions, yet remained unaltered when R81 cells were directly introduced into dye solutions. Furthermore, cells that were induced through prior cultivation in a medium containing RB19 demonstrated enhanced efficacy in decolorization compared to noninduced cells. Subsequent analysis indicated that the development of biofilms and the synthesis of polysaccharides by strain R81 were augmented in a concentration‐dependent fashion by RB19. Nevertheless, the decolorization efficacy of R81 was impeded by the existence of sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB), both of which possess the capacity to eliminate polysaccharides. The decolorization capabilities were reinstated by the SDS or CTAB eluent containing polysaccharides, suggesting a reliance on the presence of polysaccharides. The employment of stepwise diethylaminoethyl (DEAE)‐cellulose chromatography and decolorization experiments elucidated the importance of a specific polysaccharide in the decolorization process. This study proposes a bacteria‐derived polysaccharide as a promising remedy for treating dyeing wastewater contaminated with anthraquinones.

Investigation of the Reaction between a Homemade PEEK Oligomer and an Epoxy Prepolymer: Optimisation of Critical Parameters Using Physico-Chemical Methods.

Several researchers have examined the interest in using a thermoplastic to increase thermoset polymers' shock resistance. However, fewer studies have examined the nature of the mechanisms involved between both kinds of polymers. This was the objective of our work, which was carried out using a gradual approach. First, we describe the synthesis of a poly(ether ether ketone) oligomer (oPEEK) with hydroxyl terminations from the reaction of hydroquinone and 4,4'-difluorobenzophenone in N-methyl-2-pyrrolidone. Then, the main physicochemical properties of this oligomer were determined using different thermal analyses (i.e., differential scanning calorimetry (DSC), thermogravimetric (ATG), and thermomechanical analyses) to isolate its response alone. The chemical characterisation of this compound using conventional analytical chemistry techniques was more complex due to its insolubility. To this end, it was sulfonated, according to a well-known process, to make it soluble and enable nuclear magnetic resonance (NMR) and size exclusion chromatography (SEC) experiments. Additional information about the structural and chemical characteristics of the oligomer and its average molecular weight could thus be obtained. The synthesis of an oligoPEEK with α,ω-hydroxyl end-groups and a molecular weight of around 5070 g/mol was thus confirmed by NMR. This value was in accordance with that determined by SEC analysis. Next, the reaction of oPEEK with an epoxy prepolymer was demonstrated using DSC and dynamic rheometry. To this end, uncured mixtures of epoxy prepolymer (DGEBA) with different proportions of oPEEK (3, 5, 10 and 25%) were prepared and characterised by both techniques. Ultimately, the epoxy-oPEEK mixture was cured with isophorone diamine. Finally, topological analyses were performed by atomic force microscopy (AFM) in tapping mode to investigate the interface quality between the epoxy matrix and the oPEEK particles indirectly. No defects, such as decohesion areas, microvoids, or cracks, were observed between both systems.

Experimental Study on the Mechanism and Law of Low-Salinity Water Flooding for Enhanced Oil Recovery in Tight Sandstone Reservoirs.

Currently, research surrounding low-salinity water flooding predominantly focuses on medium- to high-permeability sandstone reservoirs. Nevertheless, further investigation is necessary to implement this technique with regard to tight sandstone reservoirs. The present study comprises a series of experiments conducted on the crude oil and core of the Ordos Chang 6 reservoir to investigate the influence of ionic composition on low-salinity water flooding in tight oil reservoirs. The change in wettability on the rock surface was analyzed by using the contact angle experiment. The change in recovery rate was analyzed using a core displacement experiment. The reaction between rock fluids was analyzed using an ion chromatography experiment. Additionally, a nuclear magnetic resonance (NMR) experiment was used to analyze the mobilization law of crude oil and the change in wettability on the scale of the rock core. This led to a comprehensive discussion of the law and mechanism of enhancing the recovery rate via low-salinity water flooding from various perspectives. Experiments show that low-salinity water flooding is an effective technique for enhancing recovery in tight sandstone reservoirs. Altering the ionic composition of injected water can improve the water wettability of the rock surface and enhance recovery. Decreasing the mass concentration of Ca2+ or increasing the mass concentration of SO42- can prompt the ion-exchange reaction on the rock surface and detachment of polar components from the surface. Consequently, the wettability of the rock surface strengthens, augmenting the recovery process. Nuclear magnetic resonance experiments evidence that low-salinity water injection, with ion adjustment, significantly alters the interactions between the rock and fluid in tight sandstone reservoirs. As a result, the T2 signal amplitude decreases significantly, residual oil saturation reduces considerably, and the hydrophilic nature of the rock surface increases.

Understanding electrostatic interaction on strong cation-exchanger via co-ion valency effects

Empirical adsorption models have been extensively used to design and optimize ion exchange chromatography (IEC) processes for proteins. The equations go 40 years back to the qualitative findings about the electrical double layer (EDL) in ion exchangers and form the basis of the stoichiometric displacement (SD) model widely used in preparative chromatography. While the SD model reduces the experimental effort to find salt-eluting conditions for the separation, knowledge transfer is restricted from one system to another. However, this limitation can be overcome by understanding the physicochemical interaction mechanism between the solid adsorbent and the electrolyte. Via a theoretical and experimental approach, we investigated the physicochemical adsorption mechanism in IEC and developed a methodology to determine it quantitatively by measuring the effective EDL thickness. We performed negative adsorption experiments in high-performance liquid chromatography to measure the excluded volume of co-ions, citrate, or oxalate on strong cation exchange resin. Together with the physical specifications of the column and the deployment of a modified nonlinear Poisson-Boltzmann equation, we identified the effects of the electrolyte composition on the size of the EDL. While it depends on the concentration, valency, and size of the counterion, we derived that the expansion of the EDL is indicated by different valencies of the carboxylate co-ions in trace amounts. Our findings provide a self-consistent theory of the transport phenomena in a solid/fluid system with all parameters specified with the physical properties of the chromatographic process. Further, optimizing the resin design or improving the adsorption and desorption conditions for biomolecules may be facilitated. Altogether, our work may improve material designing and process development and, thereby, help to overcome the concurrent technological and economic bottlenecks of the well-deployed purification step of IEC.

Hydrophobic Components in Light-Yellow Pulp Sweet Potato (Ipomoea batatas (L.) Lam.) Tubers Suppress LPS-Induced Inflammatory Responses in RAW264.7 Cells via Activation of the Nrf2 Pathway.

Sweet potato is a crop that is widely consumed all over the world and is thought to contribute to health maintenance due to its abundant nutrients and phytochemicals. Previous studies on the functionality of sweet potatoes have focused on varieties that have colored pulp, such as purple and orange, which contain high levels of specific phytochemicals. Therefore, in the present study, we evaluated the anti-inflammatory effects of light-yellow-fleshed sweet potatoes, which have received little attention. After freeze-drying sweet potatoes harvested in 2020, extracts were prepared from the leaves, stems, roots, and tubers in 100% ethanol. Mouse macrophage-like cell line RAW264.7 cells were cultured with 10 µg/mL of the extracts and induced lipopolysaccharide (LPS)-stimulated inflammation. Of the extracts, the tuber extracts showed the highest suppression of LPS-induced interleukin-6 (IL-6) gene expression and production in RAW264.7, which was attributed to the activation of the nuclear factor erythroid 2-related factor 2 (Nrf2) oxidative stress response pathway. In addition, preparative high-performance liquid chromatography (HPLC) experiments suggested that hydrophobic components specific to the tuber were the main body of activity. In previous studies, it has been shown that the tubers and leaves of sweet potatoes with colored pulp exhibit anti-inflammatory effects due to their rich phytochemicals, and our results show that the tubers with light-yellow pulp also exhibit the effects. Furthermore, we were able to show a part of the mechanism, which may contribute to the fundamental understanding of the treatment and prevention of inflammation by food-derived components.

Identification and Toxicity Profiling of Column Fractions of Ethanol Leaf Extract of Ziziphus mauritiana

Different plants may contain varying amounts of phytochemicals and also different degrees of toxicity. This study aimed to evaluate the phytochemicals present in the most active column fraction of ethanol leaf extract of the Ziziphus mauritiana plant and their toxic effects using brine shrimp lethality assay and animal model. Crude ethanol extract was obtained by maceration while fraction was achieved using a column chromatography experiment. Toxicity was evaluated with brine shrimp lethality assay and albino rat’s models while characterization was achieved with liquid chromatography-mass spectrophotometry. Ziziphus mauritiana leaves revealed 42 fractions pooled into seven fractions. Fraction three (3) was the most toxic with the brine shrimp lethality assay of (31.48ug/ml) and its toxicological evaluation revealed an adverse effect on the hematological parameter, biochemical indices, and histo-architecture of the liver and kidney of the experimental model studied. LCMS analysis of the most toxic fraction revealed the presence of Antirrhinoside, Lucidumol A, Apigenin 7-glucuronide-4'-(6''-malonylglucoside), Dioscoreside C, Camellioside D, and others which have been reported for various pharmacological effects including adverse effects, The mode of toxicity may be synergistic, individual, or antagonistic which may explain the moderate toxicity observed in animal model. Administration of these fractions may lead to toxicity despite their potential.

Revealing the pyrolysis mechanism of polypropylene cable insulation: A combined study of TG‐GC experiments and RMD/DFT simulations

AbstractStyrene grafted polypropylene (PP‐g‐St) is one of the insulating materials for high voltage power cables. However, the process of pyrolysis gas generation in polypropylene cable insulation remains poorly understood. To address this knowledge gap, this study employed a combination of thermogravimetric‐gas chromatography experiments, density functional theory, and reactive molecular dynamics simulations. The experimental findings revealed that the pyrolysis gases primarily consisted of H2, CO, C2H4, and CH4. Higher temperatures were found to increase the yields of H2 and CO. The simulation results indicated that H2 and CO were generated through the rupture of less reactive olefins and radicals, while C2H4 was primarily produced by the rupture of CC bonds in the polypropylene chain. Additionally, CH4 was formed when CH3 groups captured hydrogen from other molecules. By the chain reaction mechanism, we enable the calculation of the activation energy of PP‐g‐St. This study provides a theoretical foundation for understanding the pyrolysis gas generation.

Study on the pyrolysis characteristics of styrene-grafted polypropylene cable insulation material

Polypropylene (PP) has garnered significant attention as a cable insulation material due to its exceptional electrical performance and recyclability. Styrene-grafted polypropylene (PP-g) emerges as a promising alternative in this context. This study employs thermogravimetric-gas chromatography experiments and reactive molecular dynamics simulations to compare the pyrolysis process and decomposition products of pure PP and PP-g as cable insulation materials. Results indicate that while both materials produce similar pyrolysis products, PP-g exhibits greater resistance to decomposition than PP. The predominant hydrocarbons in the decomposition products include methane (CH4), ethylene (C2H4), propylene (C3H6), and ethane (C2H6). The relative proportion of hydrocarbons decreases with increasing temperature, with a shift toward increased C2H6 production. The ratio of C2H6 to (CH4 + C2H4) molecular weights in thermal decomposition products rises with temperature, suggesting enhanced stability. Graft modification alters the decomposition pathway, specifically increasing the generation of C2H6. This study lays a theoretical foundation for cable aging monitoring and life assessment.

Parameter estimation for reactive chromatography model by Bayesian inference and parallel sequential Monte Carlo

Reactive chromatography overcomes many disadvantages of conventional separation processes by improving yields and reduces costs by combining reaction and separation in a single operation. Model-based optimization is essential to realize industrial applications of this process. Parameters in the model must be estimated accurately, and evaluating the uncertainty of the model parameters is crucial. However, uncertainty quantification of model parameters has not been performed for reactive chromatography processes. In this study, we propose an approach to estimate the parameter uncertainty of reactive chromatography processes using Bayesian inference and parallel sequential Monte Carlo. As an example, the esterification synthesis of acetic acid and methanol catalyzed by a cation exchange resin was considered. Parameter estimation was performed using a reactive chromatography experiment and a non-reactive experiment in which only the products were injected. The results of the analysis showed that using both the reactive and non-reactive chromatography experimental data simultaneously improved the accuracy of the estimation. In addition, correlations between some parameters were revealed.

Factors affecting product association as a mechanism of host-cell protein persistence in bioprocessing.

Product association of host-cell proteins (HCPs) to monoclonal antibodies (mAbs) is widely regarded as a mechanism that can enable HCP persistence through multiple purification steps and even into the final drug substance. Discussion of this mechanism often implies that the existence or extent of persistence is directly related to the strength of binding but actual measurements of the binding affinity of such interactions remain sparse. Two separate avenues of investigation of HCP-mAb binding are reported here. One is the measurement of the affinity of binding of individual, commonly persistent Chinese hamster ovary (CHO) HCPs to each of a set of mAbs, and the other uses quantitative proteomic measurements to assess binding of HCPs in a null CHO harvested cell culture fluid (HCCF) to mAbs produced in the same cell line. The individual HCP measurements show that the binding affinities of individual HCPs to different mAbs can vary appreciably but are rarely very high, with only weak pH dependence. The measurements on the null HCCF allow estimation of individual HCP-mAb affinities; these are typically weaker than those seen in affinity measurements on isolated HCPs. Instead, the extent of binding appears correlated with the initial abundance of individual HCPs in the HCCF and the forms of the HCPs in the solution, i.e., whether HCPs are present as free molecules or as parts of large aggregates. Separate protein A chromatography experiments performed by feeding different fractions of a mAb-containing HCCF obtained by size-exclusion chromatography (SEC) showed clear differences in the number and identity of HCPs found in the protein A eluate. These results indicate a significant role for HCP-mAb association in determining HCP persistence through protein A chromatography, presumably through binding of HCP-mAb complexes to the resin. Overall, the results illustrate the importance of considering more fully the biophysical context of HCP-product association in assessing the factors that may affect the phenomenon and determine its implications. Knowledge of the abundances and the forms of individual or aggregated HCPs in HCCF are particularly significant, emphasizing the integration of upstream and downstream bioprocessing and the importance of understanding the collective properties of HCPs in addition to just the biophysical properties of individual HCPs.

Leaching of plasticizers from PVC medical devices: A molecular interpretation of experimental migration data

High pressure liquid chromatography (HPLC) experiments and molecular simulations were combined to investigate the leaching process of DINCH, DEHT and TOTM plasticizers from PVC materials. Methodologies based on statistical thermodynamics were used to rationalize the migration of the different plasticizers in water and ethanolic solutions from different free energy contributions. We extended this combined approach to the study of the adsorption of a model drug (diazepam) on different plasticized-PVC materials. The drug loss by sorption was investigated in different solutions and rationalized at an energy level. The combination of experiments and theoretical approaches proved to be effective for these sophisticated systems and processes for which various energy contributions are involved.

Accelerating Pharmaceutical Process Development with an Acoustic Droplet Ejection-Multiple Reaction Monitoring-Mass Spectrometry Workflow.

Fast-paced pharmaceutical process developments (e.g., high-throughput experimentation, directed evolution, and machine learning) involve the introduction of fast, sensitive, and accurate analytical assays using limited sample volumes. In recent years, acoustic droplet ejection (ADE) coupled with an open port interface has been invented as a sampling technology for mass spectrometry, providing high-throughput nanoliter analytical measurements directly from the standard microplates. Herein, we introduce an ADE-multiple reaction monitoring-mass spectrometry (ADE-MRM-MS) workflow to accelerate pharmaceutical process research and development (PR&D). This systematic workflow outlines the selection of MRM transitions and optimization of assay parameters in a data-driven manner using rapid measurements (1 sample/s). The synergy between ADE sampling and MRM analysis enables analytical assays with excellent sensitivity, selectivity, and speed for PR&D reaction screenings. This workflow was utilized to develop new ADE-MRM-MS assays guiding a variety of industrial processes, including (1) screening of Ni-based catalysts for C-N cross-coupling reaction at 1 Hz and (2) high-throughput regioisomer analysis-enabled enzyme library screening for peptide ligation reaction. ADE-MRM-MS assays were demonstrated to deliver accurate results that are comparable to conventional liquid chromatography (LC) experiments while providing >100-fold throughput enhancement.

Application of Comparative Teaching Method in Experimental Project Design of Instrumental Analysis Course: A Case Study in Chromatography Experiment Teaching

Application of Comparative Teaching Method in Experimental Project Design of Instrumental Analysis Course: A Case Study in Chromatography Experiment Teaching

Reactivity of Ts and At oxides and oxyhydrides with a gold surface from periodic DFT calculations.

Adsorption energies, Eads, of oxides and oxyhydrides of the superheavy element (SHEs) Ts and of its lighter homologue At on the gold surface are predicted on the basis of relativistic periodic density functional theory calculations via AMS BAND software. The following compounds were considered: MO, MO2, MOO, and MO(OH) (where M = At and Ts). The aim of this study is to support "one-atom-at-a-time" gas-phase chromatography experiments on reactivity/volatility of SHEs. The results obtained indicate that all the molecules investigated should interact fairly strongly with the gold surface, with those of Ts being more reactive than At ones. The similarity in the Eads values of all the considered At compounds would make it challenging to differentiate between them while measuring their adsorption enthalpies, given experimental uncertainty. However, the difference in Eads among Ts compounds is more pronounced, so that one should be able to differentiate between the species.

Selective Transesterification to Control Copolymer Microstructure in the Ring-Opening Copolymerization of Lactide and ε-Caprolactone by Lanthanum Complexes.

A series of novel lanthanum amido complexes, supported by ligands designed around the salan framework (salan = N,N'-bis(o-hydroxy, m-di-tert-butylbenzyl)-1,2-diaminoethane) were synthesized and fully characterized in the solid and solution states. The ligands incorporate benzyl or 2-pyridyl substituents at each tertiary amine center. The complexes were investigated as catalysts in the ring-opening homopolymerization of lactide (LA) and ε-caprolactone (ε-CL) and copolymerization of equimolar amounts of LA and ε-CL at ambient temperature. Solvent (THF or toluene) and the number of 2-pyridyl groups in the complex were found to influence the reactivity of the catalysts in copolymerization reactions. In all cases, complete conversion of LA to PLA was observed. The use of THF, a coordinating solvent, suppressed ε-CL polymerization, while the presence of one or more 2-pyridyl groups promoted ε-CL polymerization. Each copolymer gave a monomodal trace in gel permeation chromatography-size-exclusion chromatography (GPC-SEC) experiments, indicative of copolymer formation over homopolymerization. Copolymer microstructure was found to be dependent on catalyst structure and reaction solvent, ranging from blocky to close to alternating. Experiments revealed rapid conversion of LA in the initial stages of the reaction, followed by incorporation of ε-CL into the copolymer by either transesterification or propagation reactions. Significantly, the mode of transesterification (TI or TII) that occurs is determined by the structure of the metal complex and the reaction solvent, leading to the possibility of controlling copolymer microstructure through catalyst design.

THE INFLUENCE OF PENICILLIUM FUNGI ISOLATED FROM SOYBEAN ROOTS ON THE SYMBIOTIC SYSTEM “GLYCINE MAX – BRADYRHIZOBIUM JAPONICUM” AND CROP PRODUCTIVITY

Objective. Identify new strains of Penicillium endophytic fungi by morphological and cultural properties, study their influence on the symbiotic system “Glycine max – Bradyrhizobium japonicum” and soybean productivity. Methods. Microbiological (cultivation of endophytic fungi, preparation of spore suspensions, determination of spore titre in suspensions, light microscopy), gas chromatographic (determination of nitrogenase activity of soybean nodules), vegetation and field small-plot experiments (study of the effect of endophytic fungi on symbiotic and morphometric parameters of soybean plants and crop yield when growing on sod-podzolic soil), statistical. Results. New strains of Penicillium endophytic fungi are attributed to the species P. funiculosum Thom, P. variabile Sopp, P. glauco-lanosum Chalabuda, and P. steckii Zaleski based on their morphological and cultural characteristics. Under the conditions of the vegetation experiment, the positive influence of the studied strains on the symbiotic parameters of soybeans was shown. A tendency towards an increase in the number of nodules versus the control in all variants, an increase in the mass of nodules in the variants when soybean seeds were treated with spores of P. variabile 20173 and P. funiculosum 20312 by 28.4 % and 36.4 %, respectively, an increase in the nitrogenase activity of nodules by 29.8–38.5 % in variants with the use of P. stekii 2204, P. glauco-lanosum 20401, P. variabile 20173 was registered. The results of the study of soybean symbiotic parameters under the conditions of the field experiment generally confirmed the corresponding data of the vegetation experiment. Inoculation of soybean seeds with spores of Penicillium endophytic fungi contributed to an increase of the morphometric parameters of plants, namely: height — by 4.6–6.2 cm in all variants of the experiment, above-ground mass of dried plants — by 11.1–19.0 % when treated with P. stekii 2204, P. glauco-lanosum 20401, P. variabile 20173, mass of dry roots — by 13.2 % and 18.4 % with P. funiculosum 20312 and P. variabile 20173. The results of the field experiment proved the positive influence of new strains of endophytic fungi on soybean yield: when soybean seeds were inoculated with P. variabile 20173, P. funiculosum 20312 and P. stekii 2204, the increase in grain productivity was on average from 11.8 % to 17.3 %. The weight of 1000 grains increased by 3.8–5.1 % in variants using P. variabile 20173, P. glauco-lanosum 20401, P. stekii 2204. Conclusion. Penicillium endophytic fungi, which belong to the species P. funiculosum, P. variabile, P. glauco-lanosum and P. stekii, were isolated from the tissues of soybean roots. It was found that Penicillium endophytic fungi have a positive influence on the functioning of the symbiotic system “Glycine max – Bradyrhizobium japonicum” and soybean yield.

Tanc1/2 TPR domain interacts with Myo18a C-terminus and undergoes liquid-liquid phase separation

Tanc1 and its homologous protein Tanc2 are critical synaptic scaffold proteins which regulate synaptic spine densities and excitatory synapse strength. Recent studies indicated TANC1 and TANC2 are candidate genes of several neurodevelopmental disorders (NDDs). In this study, we identified and characterized a novel interaction between Tanc1/2 and Myo18a, mediated by the Tanc1/2 TPR domains and Myo18a coiled-coil domain and C-extension (CCex). Sequence analysis and size exclusion chromatography experiments reveal that high salt disrupts the interaction between Myo18a and Tanc1/2, indicating that the interaction is primarily driven by charge-charge interactions. More importantly, we found that the Tanc1-TPR/Myo18a CCex interaction could undergo liquid-liquid phase separation (LLPS) in both cultured cells and test tubes, which provides the biochemical basis and potential molecular mechanisms for the LLPS-mediated interactions between Myo18a and Tanc1/2.

Exploring the Effect of Compound Glycyrrhizin and Silybinin on the Metabolism of Pexidartinib in Rats Based on CYP3A4 and CYP2C9

Pexidartinib offered a new therapeutic option for adult patients with symptomatic tenosynovial giant cell tumor (TGCT) who were refractory to surgical treatment and had severe morbidity or functional limitations. Meanwhile, the metabolism of pexidartinib was mainly mediated through the oxidation of cytochrome P450 (CYP) 3A and glucuronidation by uridine glucuronosyltransferase (UGT) 1A4 and attention shall be paid to CYP450-based drug-drug interactions during therapeutic dosing. This study aimed to examine the changes in the pharmacokinetics of pexidartinib by silymarin and compound glycyrrhizin on pexidartinib in vivo in rats by high-performance liquid chromatography (HPLC)-UV approach and to detect its expression in CYP3A4 and CYP2C9 using the western blot. The findings of chromatography experiments revealed that silybinin as well as compound glycyrrhizin increased the exposure of pexidartinib in rats and had a significant inhibitory effect on the metabolism of pexidartinib. The results of immunoblotting assays suggested that silybinin as well as compound glycyrrhizin inhibited the protein expression of CYP3A4 and CYP2C9 in rats. Therefore, the combination of pexidartinib with silybinin and compound glycyrrhizin should be monitored to avoid clinical adverse effects.

Experimental Study on the Control Mechanism of Non-Equilibrium Retrograde Condensation in Buried Hill Fractured Condensate Gas Reservoirs

During the depletion development of condensate gas reservoirs, when the formation pressure drops below the dew point pressure, the condensate oil and natural gas systems are in the non-equilibrium state of foggy retrograde condensation. The rational use of the non-equilibrium phase characteristics of the foggy retrograde condensation phenomenon during the development process will be beneficial to the recovery of condensate oil and natural gas. In order to clarify the retrograde condensation control mechanism during the non-equilibrium depletion development of condensate gas reservoirs, the phase characteristics of a condensate oil and gas system were studied by constant composition expansion and constant volume depletion experiments. Then, on the basis of a long core depletion experiment and chromatographic analysis experiment, the influence of different pressure drop speeds, fluid properties, and reservoir physical properties on the control effect of non-equilibrium retrograde condensation after the coupling of the fluid retrograde condensation and reservoir core is analyzed. The results show that during the pressure decline process, the condensate oil and gas system will produce a strong foggy retrograde condensation phenomenon, with the saturation of the retrograde condensate increasing and then decreasing. The cumulative recovery of the condensate oil and natural gas, as well as the mass fraction of the heavy components in the condensate oil, increase with the increase in the depletion rate. Different fluid properties and reservoir physical properties have a great influence on the cumulative recovery degree of the condensate oil, and have little influence on the recovery degree of the natural gas. This work has a certain guiding role for the stable production and enhanced recovery of fractured condensate gas reservoirs in subsurface structures of metamorphic rocks.

Establishment of a quantitative method for GC analysis of polyoxyethylene (35) castor oil in microemulsion extracts

With the continuous exploration of microemulsions as solvents for traditional Chinese medicine extraction, polyoxyethy-lene(35) castor oil(CrEL), a commonly used surfactant, is being utilized by researchers. However, the problem of detecting residues of this surfactant in microemulsion extracts has greatly hampered the further development of microemulsion solvents. Based on the chemical structures of the components in CrEL and the content determination method of castor oil in the 2020 edition of the Chinese Pharmacopoeia(Vol. Ⅳ), this study employed gas chromatography(GC) and single-factor experiments to optimize the preparation method of methyl ricinoleate from CrEL. The conversion coefficient between the two was validated, and the optimal sample preparation method was used to process microemulsion extracts of Zexie Decoction from three batches. The content of methyl ricinoleate generated was determined, and the content of CrEL in the microemulsion extracts of Zexie Decoction was calculated using the above conversion coefficient. The results showed that the optimal preparation method for CrEL was determined. Specifically, 10 mL of 1 mol·L~(-1) KOH-methanol solution was heated at 60 ℃ for 15 min in a water bath. Subsequently, 10 mL of boron trifluoride etherate-methanol(1∶3) solution was heated at 60 ℃ for 15 min in a water bath, followed by extraction with n-hexane twice. CrEL could stably produce 20.84% methyl ricinoleate. According to this conversion coefficient, the average mass concentration of CrEL in the three batches of Zexie Decoction microemulsion extracts was 11.94 mg·mL~(-1), which was not significantly different from the CrEL mass concentration of 11.57 mg·mL~(-1) during microemulsion formulation, indicating that the established content determination method of this study was highly accurate, sensitive, and repeatable. It can be used for subsequent research on microemulsion extracts of Zexie Decoction and provide a reference for quality control of other drug formulations containing CrEL.