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We established an efficient method using high-speed countercurrent chromatography (HSCCC) combined with preparative high-performance liquid chromatography (prep-HPLC) for isolating and purifying Gelsemium elegans (G. elegans) alkaloids. First, the two-phase solvent system composed of 1% triethylamine aqueous solution/n-hexane/ethyl acetate/ethanol (volume ratio 4:2:3:2) was employed to separate the crude extract (350 mg) using HSCCC. Subsequently, the mixture that resulted from HSCCC was further separated by Prep-HPLC, resulting in seven pure compounds including: 14-hydroxygelsenicine (1, 12.1 mg), sempervirine (2, 20.8 mg), 19-(R)-hydroxydihydrogelelsevirine (3, 10.1 mg), koumine (4, 50.5 mg), gelsemine (5, 32.2 mg), gelselvirine (6, 50.5 mg), and 11-hydroxyhumanmantenine (7, 12.5 mg). The purity of these seven compounds were 97.4, 98.9, 98.5, 99, 99.5, 96.8, and 85.5%, as determined by HPLC. The chemical structures of the seven compounds were analyzed and confirmed by electrospray ionization mass spectrometry (ESI-MS), 1H-nuclear magnetic resonance (1H NMR), and 13 C-nuclear magnetic resonance (13 C NMR) spectra. The results indicate that the HSCCC-prep-HPLC method can effectively separate the major alkaloids from the purified G. elegans, holding promising prospects for potential applications in the separation and identification of other traditional Chinese medicines. [ABSTRACT FROM AUTHOR]

Athenaea fasciculata belongs to the Solanaceae family and is a promising source of cytotoxic withanolides known as aurelianolides A and B. In the last years, the pharmacological studies of these aurelianolides on different leukemia cell lines have stimulated new studies on their potential as alternative candidates for new lead anticancer drugs. However, the obtention of these two pure compounds by traditional preparative is a costly and long time-consuming process, which is performed in several steps. This study aimed to propose a straightforward approach for isolating aurelianolides A and B using high-speed countercurrent chromatography (HSCCC). In this study, among 10 different solvent systems, the system composed of n-hexane/ethyl acetate/methanol/water 3:6:2:1 (v/v/v/v) was chosen for optimization. This HEMWat system was optimized to 4:8:2:4 (v/v/v/v) and chosen for HSCCC separation in a tail-to-head elution mode. After the HSCCC scale-up procedure, a withanolides mixture (200.0 mg) was separated within 160 min in a single-step purification process. In total, 78.9 mg of aurelianolide A (up to 95.0% purity) and 54.3 mg of aurelianolide B (up to 88.5% purity) was obtained by this fast sequential liquid–liquid partition process. The isolated withanolides were identified by 1H and 13C NMR spectroscopy (this method has proven to be faster and more efficient than classical procedures (CC and Prep-TLC)). [ABSTRACT FROM AUTHOR]

AbstractIn this study, biocatalytic transesterification reaction using Novozyme 435® (N435) lipase was employed to enhance the hydrophobicity of esculin, aiming to improve its solubility for commercial applications and enhance its bioactivity and oral viability. The acylation reaction of esculin with vinyl acetate was conducted at 60 °C and 200 rpm for 24 h. After chromatographic and spectroscopic analysis, two products were identified: the first one was monoacylated at the 6′-OH position of the glucosyl moiety of esculin (TR: 10.3 min and m/z 382.93 [M + H]+), and the second one was diacylated at the 6′-OH and 3′-OH positions (TR: 13.0 min and m/z 424.93 [M + H]+). The latter was the major product, with a conversion rate of 53.550 ± 0.368%, while the monoacetylated one showed 8.715 ± 0.064%. Both products were isolated by high-speed counter-current chromatography (HSCCC) using a two-phase system HEMWat 1:9:1:9 and characterized by NMR. In this way, these results improve the practical application of esculin, through the obtention of esculin mono and diacetates by fast and efficient biocatalysis reaction. [ABSTRACT FROM AUTHOR]

Acetylcholinesterase inhibitors from Evodia rutaecarpa were screened, prepared, and evaluated. To screen the lipophilic alkaloid active constituents in E. rutaecarpa, we improved and optimized an ultrafiltration system. Three acetylcholinesterase (AChE) inhibitors, dehydroevodiamine, evodiamine, and rutecarpine, were screened. Addressing the limitations of the traditional response surface methodology (RSM) for multiobjective screening, we integrated RSM with the Non‐dominated Sorting Genetic Algorithm III to achieve the optimal extraction of these active ingredients. High‐speed countercurrent chromatography was used to isolate the active components using a two‐phase solvent system: n‐hexane/ethyl acetate/methanol/water (3.0:2.5:3.5:2.0, v/v/v/v) and ethyl acetate/methanol/water (3.0:1.0:4.0, v/v/v). The nuclear magnetic resonance spectroscopy confirmed the structures of the compounds, and molecular docking and dynamics simulations assessed the inhibitory effects of the chemical components on AChE, which were consistent with the findings of the ultrafiltration experiments. We also confirmed the neuroprotective properties of these compounds against glutamate‐induced apoptosis in PC12 cells. Overall, we achieved the systematic optimization of multitarget compound extraction and lipophilic alkaloid ultrafiltration screening, as well as preparation and activity validation, laying the groundwork for the development of AChE inhibitors from lipophilic alkaloids. [ABSTRACT FROM AUTHOR]

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The essential oil components of traditional Chinese medicine in-hospital preparation were complex, and one-dimensional chromatographic separation was difficult to completely separate them due to the limited peak capacity. This study was carried out to establish a comprehensive two-dimensional chromatographic separation and analysis method based on countercurrent chromatography (CCC) and gas chromatography (GC). In this paper, we focused on the separation of the essential oil of the traditional Chinese medicine in-hospital preparation Wenjing Tongluo San by CCC × GC, and explored the orthogonality between the two chromatographic techniques to provide the new technical support for the screening of the active ingredients. A solvent system composed of n-hexane-ethyl acetate-methanol-water (9.5:0.5:8.5:1.5, v/v) was chosen for the first-dimensional CCC separation. All the fractions collected from CCC were transferred to GC for plotting two-dimensional contours map. The calculated capacity of the two-dimensional separation system exceeded 3000, which was 8 times more than that of the one-dimensional separation system. High orthogonality (r = 0.42) and spatial coverage factor (70.42%) were obtained. Meanwhile, all the fractions were identified by GC-MS. Our research provided a new methodology for separating essential oils in traditional Chinese medicine as well as an approach for evaluating the quality of traditional Chinese medicinal in-hospital preparation based on two-dimensional chromatographic fingerprints. [ABSTRACT FROM AUTHOR]

Owing to its ability to separate substances with a broad scope of polarities, exploring the three‐phase solvent systems (TPSSs) with high‐speed countercurrent chromatography is a topic of interest in separation science, and their retention volumes should be more concerned. This study primarily investigates the behavior of retention volumes while examining the isolation abilities of the TPSS in the technique above. We took standard compounds, including sophoricoside, Sudan red 7B, and rotenone, which have a broad range of polarity, for investigation in this study and separated them using different four‐liquid TPSSs made up of water, acetonitrile, methyl acetate, and n‐hexane (WAMH). Our findings show that the retention volumes gradually alter in response to changes in phase polarity within the proposed solvent systems. With TPSSs, we preliminarily studied compound isolation and the promising formula of their retention volumes. The proposed solvent systems WAMH in different ratios showed high correlations and adjusted correlation coefficients above 0.9978 and 0.9913 for the actual and calculated retention volumes. This study will be particularly beneficial for researchers focusing on countercurrent chromatography with TPSSs, as it offers valuable time‐saving insights. [ABSTRACT FROM AUTHOR]

A continuous purification process can be beneficial to the purification of biologics due to its higher productivity and efficiency than a conventional batch purification process. However, regulatory issues and lack of established cases render deployment of the continuous process difficult in industrial settings. Here we report a case study for design and optimization of an advanced continuous process for purifying a low-titer enzyme as a model biologic. To convert a conventional batch process to an advanced continuous one in purification of biologics, conventional unit operations (UOs), including ultrafiltration/diafiltration (UF/DF) and batch chromatography, were replaced by advanced ones such as in-line dilution conditioning (IDC) and periodic counter-current chromatography (PCC). The UF/DF UO was changed to IDC UO to adjust pH and conductivity. The mixing ratio of the sample and the conditioning buffer in IDC was determined by experiments with three buffers. PCC was optimized with two variables, column height and sample loading residence time, as the delta pressure in the columns was less than 1.0 bar. A graph indicating the operating area was plotted to efficiently control the PCC. Although the sample volume increased in IDC, PCC had a complementary advantage in that purification was performed faster than batch chromatography. We observed at least 25% increase in economic advantage when the advanced continuous process was applied to purify a low-titer enzyme. We propose not only a continuous process with the substitution of UF/DF and batch chromatography with IDC and PCC but also a method to optimize PCC by plotting operating areas. [ABSTRACT FROM AUTHOR]

Four monodisperse porous polymer microspheres were successfully prepared by seed emulsion polymerization and used as stationary phases for HPLC and preparative high-performance liquid chromatography (Prep-HPLC). All four polymer microspheres(polystyrene–polystyrene (PS-PS), polystyrene-poly(glycidyl methylate) (PS-PGMA), polystyrene-poly(methyl methylate) and poly(glycidyl methylate)–poly(glycidyl methylate) were used for filling HPLC empty columns. According to the analysis results of the HPLC column, PS-PS and PS-PGMA microspheres were screened out as the stationary phase of Prep-HPLC. The industrial-grade phytol was successfully separated and purified, and the purity of the final phytol was as high as 99%. The two types of polymer microspheres have been applied to industrial-grade phytol purification and have been used in factories. [ABSTRACT FROM AUTHOR]

In this study, we employed a combination approach for the preparative separation of constituents from Ginkgo biloba L. leaves. It involved multi‐stage solvent extractions utilizing two‐phase multi‐solvent systems and countercurrent chromatography (CCC) separations using three different solvent systems. The n‐heptane/ethyl acetate/water (1:1:2, v/v) and n‐heptane/ethyl acetate/methanol/water (HepEMWat, 7:3:7:3, v/v) solvent systems were screened out as extraction systems. The polarities of the upper and lower phases in the multi‐solvent systems were adjustable, enabling the effectively segmented separation of complex constituents in G. biloba L. The segmented products were subsequently directly utilized as samples and separated using CCC with the solvent systems acetate/n‐butanol/water (4:1:5, v/v), HepEMWat (5:5:5:5, v/v), and HepEMWat (9:1:9:1, v/v), respectively. As a result, a total of 11 compounds were successfully isolated and identified from a 2 g methanol extract of G. biloba L through two‐stage extraction and three CCC separation processes; among them, nine compounds exhibited high‐performance liquid chromatography purity exceeding 85%. [ABSTRACT FROM AUTHOR]

Propolis is a resinous bee product with a very complex composition, which is dependent upon the plant sources that bees visit. Due to the promising antimicrobial activities of red Brazilian propolis, it is paramount to identify the compounds responsible for it, which, in most of the cases, are not commercially available. The aim of this study was to develop a quick and clean preparative-scale methodology for preparing fractions of red propolis directly from a complex crude ethanol extract by combining the extractive capacity of counter-current chromatography (CCC) with preparative HPLC. The CCC method development included step gradient elution for the removal of waxes (which can bind to and block HPLC columns), sample injection in a single solvent to improve stationary phase stability, and a change in the mobile phase flow pattern, resulting in the loading of 2.5 g of the Brazilian red propolis crude extract on a 912.5 mL Midi CCC column. Three compounds were subsequently isolated from the concentrated fractions by preparative HPLC and identified by NMR and high-resolution MS: red pigment, retusapurpurin A; the isoflavan 3(R)-7-O-methylvestitol; and the prenylated benzophenone isomers xanthochymol/isoxanthochymol. These compounds are markers of red propolis that contribute to its therapeutic properties, and the amount isolated allows for further biological activities testing and for their use as chromatographic standards. [ABSTRACT FROM AUTHOR]

An efficient method for the continuous separation of Voriconazole enantiomers was developed using sulfobutyl ether‐β‐cyclodextrin (SBE‐β‐CD) as a chiral selector in high‐speed countercurrent chromatography (HSCCC) with different types. The separation was performed using a two‐phase solvent system consisting of n‐hexane/ethyl acetate/100 mmol/L phosphate buffer solution (pH = 3.0, containing 50 mmol/L SBE‐β‐CD) (1.5:0.5:2, v/v/v). A fast and predictable scale‐up process was achieved using an analytical DE HSCCC instrument. The optimized parameters were subsequently applied to a preparative Tauto HSCCC instrument, resulting in consistent separation time and enantiomeric purity, with throughput boosted by a remarkable 11‐fold. Preparative HSCCC successfully separated 506 mg of the racemate, delivering enantiomers exceeding 99% purity as confirmed by high‐performance liquid chromatography analysis. This investigation presents an effective methodology for forecasting the HSCCC scale‐up process and attaining continuous separation of chiral drugs. [ABSTRACT FROM AUTHOR]

Tubers of Gymnadenia conopsea (L.) R. Br. (Orchidaceae), a traditional medicine and food homologous plant, has a broad application and development prospect in the food and drug industries. Benzylester glucosides, the main effective active components in this plant, are difficult to separate due to their similar structures and high polarity. In this study, linear gradient counter‐current chromatography was used to separate benzylester glucosides and derivatives, combined with elution‐extrusion mode. The main separation parameters were optimized, including the ratio of mobile phase and sample loading. Finally, seven compounds were successfully separated, including 4‐hydroxybenzyl alcohol (1), 4‐hydroxybenzaldehyde (2), dactylorhin B (3), loroglossin (4), dactylorhin A (5), 4‐(ethoxymethyl) phenol (6), and militarine (7). The structures were analyzed by mass spectrometry and nuclear magnetic resonance spectrometry. According to our findings, the established method was an efficient approach to separate benzylester glucosides and derivatives from tubers of G. conopsea. The established strategy could be applied to purify other similar high‐polarity compounds from complex natural products. [ABSTRACT FROM AUTHOR]

The separation of cannabinoids from hemp materials is nowadays one of the most promising industrial applications of liquid‐liquid chromatography (LLC). Despite various experimental research efforts to purify cannabinoids, there are currently few works on process modeling. Thus, this study aimed to explore a straightforward approach to model the LLC separation of cannabinoids from two hemp extracts with different compositions. The feed materials were simplified to mixtures of preselected key components (i.e., cannabidiol, tetrahydrocannabinol, cannabigerol, and cannabinol). The elution profiles of cannabinoids were simulated using the equilibrium‐cell model with an empirical nonlinear correlation. The model parameters were derived from the elution profiles of single‐solute pulse injections. For the validation of the proposed approach, LLC separations with the two hemp extracts were performed in descending mode with the solvent system composed of hexane/methanol/water 10/8/2 (v/v/v). The injected sample concentrations were gradually increased from 5 to 100 mg/mL. The results showed that the approach could describe reasonably well the elution behavior of the cannabinoids, with deviations of only 1–2 min between simulated and experimental elution times. However, to improve the prediction accuracy, the model parameters can be refitted to the elution profiles of 3–4 systematically selected pulse injections with specific hemp extracts. [ABSTRACT FROM AUTHOR]

This study presents a comprehensive strategy for the selection and optimization of solvent systems in countercurrent chromatography (CCC) for the effective separation of compounds. With a focus on traditional organic solvent systems, the research introduces a "sweet space" strategy that merges intuitive understanding with mathematical accuracy, addressing the significant challenges in solvent system selection, a critical bottleneck in the widespread application of CCC. By employing a combination of volume ratios and graphical representations, including both regular and trirectangular tetrahedron models, the proposed approach facilitates a more inclusive and user‐friendly strategy for solvent system selection. This study demonstrates the potential of the proposed strategy through the successful separation of gamma‐linolenic acid, oleic acid, and linoleic acid from borage oil, highlighting the strategy's effectiveness and practical applicability in CCC separations. [ABSTRACT FROM AUTHOR]

In this study, a combination approach involving macroporous resin (MR) column chromatography and gradient countercurrent chromatography (CCC) was employed to enrich and purify bufadienolides from the roots and rhizomes of Helleborus thibetanus Franch. Initially, a D101 MR‐packed column chromatography was utilized for fractionation and enrichment of the bufadienolides, which were effectively eluted from the column using a 60% ethanol solution. CCC was subsequently introduced to separate the enriched product using the ethyl acetate/n‐butanol/water (EBuWat, 4:1:5, v/v) and EBuWat (5:0:5, v/v) solvent systems in a gradient elution mode. As results, five bufadienolides, including 6.1 mg of hellebrigenin‐3‐O‐β‐D‐glucoside (1), 2.2 mg of tigencaoside A (2), 8.3 mg of deglucohellebrin (3), 3.5 mg of 14 β‐hydroxy‐3β‐[β‐D‐glucopyranosyl‐(1→6)‐(β‐D‐glucopyranosyl)oxy]‐5α‐bufa‐20,22‐dienolide (4), and 3.0 mg of 14β‐hydroxy‐3β‐[(β‐D‐glucopyranosyl)oxy]‐5α‐bufa‐20,22‐dienolide (5), were effectively separated from 300 mg of the enriched product. The respective high‐performance liquid chromatography purities were as follows: 95.2%, 75.8%, 85.7%, 82.3%, and 92.8%. This study provides valuable insights for the efficient enrichment and separation of bufadienolides from Helleborus thibetanus Franch. [ABSTRACT FROM AUTHOR]

The article focuses on the importance of separation process technology in chemical manufacturing, highlighting its historical development and ongoing improvements in methods like distillation, extraction, adsorption, crystallization, and membrane use. It discusses the concept of separation power, particularly in distillation operations, as a key metric for evaluating and optimizing binary separation processes.

Theaflavin is a high-quality component of black tea, which has good medical and health functions. Highspeed countercurrent chromatography (HSCCC) has been applied for the separation of theaflavins and catechins, and examined the cytotoxic activity of the individual theaflavins against human liver cancer (HepG2) cells, Colonic adenocarci-noma(Caco-2)cells. Six components were separated and prepared by HSCCC. Further analysis of these compounds by HPLC indicated that that each component F4 contains theaflavin (TF), F5 contains theaflavin-3-gallate (TF-3-G) and theaflavin-3'-gallate (TF-3'-G), and F6 contains theaflavin-3, 3'- gallate (TFDG). The results show TFDG was selected to study the inhibitory effect on cancer cells. CCK-8 was used to assess cell proliferation. TFDG has a significant inhibitory effect on HepG-2 and Caco-2 cells in a time-dependent and dose-dependent manner with the increase of concentration and time. It found a solvent system for the preparation of theaflavins in large quantities was selected based on the partitioning coefficient K and separation factor a. Also, we acknowledged that there is a certain relationship between cancer prevention and quinone reductase (QR). Combining experimental data with theory, by linking molecular docking technology was used to verify that TFDG and QR can form stable complexes, It also inducing QR activity can enhance the anticancer effect, it can providing a basis for further research on the effect of theaflavin on cancer cells. Keywords high speed countercurrent chromatography; hepatoma cells; colon adenocarcinoma cells; molecular docking [ABSTRACT FROM AUTHOR]

Hericium erinaceus has long been favored for its remarkable nutritional and health-promoting benefits, and erinacine A is the key component responsible for the neuroprotective properties of H. erinaceus. Establishing an efficient method for separating erinacine A from H. erinaceus and screening the erinacine A-enriched strains is crucial to maximizing its benefits. Herein, we first reported that high-speed counter current chromatography (HSCCC) is an effective method for separating high-purity erinacine A. Using a two-phase solvent system composed of n-hexane/ethyl acetate/methanol/water (4.5:5:4.5:5, v/v/v/v), erinacine A with a purity of over 95% was separated. Then, we evaluated the content and yield of erinacine A in the liquid-fermented mycelia of Hericium germplasms. Both the content and yield of erinacine A varied greatly among the surveyed strains. The significant effect of the strain on the erinacine A content and yield was revealed by an analysis of variance. The highest erinacine A content and yield were observed in the mycelia of a wild strain HeG, reaching 42.16 mg/g and 358.78 mg/L, which is superior to the current highest outcomes achieved using submerged cultivation. The isolation method established and the strains screened in this study can be beneficial for the scaling up of erinacine A extraction and nutraceutical development to industrial levels. [ABSTRACT FROM AUTHOR]

Implantable and wearable bioelectronic systems can enable tailored therapies for the effective management of long-term diseases, thus minimising the risk of associated complications. In this context, glucose fuel cells hold great promise as in- or on-body energy harvesters for ultra-low-power bioelectronics and as self-powered glucose sensors. We report here the generation of gold nanostructures through a gold electrodeposition method in a soft template for the abiotic electrocatalysis of glucose in glucose fuel cells. Two different types of soft template were used: a lipid cubic phase-based soft template composed of Phytantriol and Brij®-56, and an emulsion-based soft template composed of hexane and sodium dodecyl sulphate (SDS). The resulting gold structures were first characterised by SAXS, SEM and TEM to elucidate their structure, and then their electrocatalytic activity towards glucose was compared in both a three-electrode set-up and in a fuel cell set-up. The Phytantriol/Brij®-56 template led to a nanofeather-like Au structure, while the hexane/SDS template led to a nanocoral-like Au structure. These templated electrodes exhibited similar electrochemical active surface areas (0.446 cm2 with a roughness factor (RF) of 14.2 for Phytantriol/Brij®-56 templated nanostructures and 0.421 cm2 with an RF of 13.4 for hexane/SDS templated nanostructures), and a sensitivity towards glucose of over 7 μA mM−1 cm−2. When tested as the anode of an abiotic glucose fuel cell (in a phosphate-buffered solution with a glucose concentration of 6 mM), a maximum power density of 7 μW cm−2 was reached; however the current density in the case of the fuel cell with the Phytantriol/Brij®-56 templated anode was approximately two times higher, reaching the value of 70 μA cm−2. Overall, this study demonstrates two simple, cost-effective and efficient strategies to manipulate the morphology of gold nanostructures, and thus their catalytic property, paving the way for the successful manufacturing of functional abiotic glucose fuel cells. [ABSTRACT FROM AUTHOR]

Carotenes, which simultaneously have the properties of natural dyes and biologically active substances that have a positive effect on human health and life expectancy, are becoming increasingly widespread in the food, pharmacological and cosmetic industries. This necessitates an increase in the production of carotenes on an industrial scale. For use in food technologies, an encapsulated form of carotenes is recommended, which requires their high purity. The purpose of this study is to analyze domestic and foreign scientific, technical and patent information on existing methods for obtaining carotenes from carotenoid concentrates to determine the direction of research for the development of technology for the production of carotenes, in particular lycopene, ensuring the production of high purity lycopene from carotenoid concentrates, as well as increasing release of lycopene. The issues of using chromatographic methods for isolating lycopene, their advantages and disadvantages, as well as the use of non-polar and low-polar organic solvents for isolating lycopene from carotenoid concentrate are considered. The promising direction of research on the use of adsorption resins in the isolation of lycopene from carotenoid concentrates is noted.It has been concluded that the currently existing methods for obtaining carotenes from carotenoid concentrates, in particular lycopene, are quite technically complex and time-consuming. Some of the mallow the production of high purity lycopene, but the yield of lycopene is so low that their industrial scaling seems impractical. Taking this into account, research is being updated in the field of developing a technology for producing lycopene from a carotenoid concentrate, providing a high yield of high purity lycopene, with the possibility of its implementation on an industrial scale.

Reductive catalytic fractionation (RCF) is a lignin-first biorefining technique that produces a polysaccharide-rich pulp and a lignin oil that is rich in aromatic monomers from aryl–ether bond cleavage and carbon–carbon linked aromatic oligomers. Separations of the lignin-derived monomers, both from one another and from the oligomers, out of these lignin oils could potentially yield high value co-products. To that end, we demonstrate that counter-current chromatography (CCC) is an effective means for simultaneous lignin monomer–monomer and monomer–oligomer separations using oils from RCF reactions with hardwood, softwood, and herbaceous feedstocks. Partition coefficient measurements of aromatic monomers from RCF of poplar, pine, and corn stover were first used to inform CCC solvent selection. We subsequently demonstrated CCC separations of those lignin oils using the HEMWat −3 solvent system and refined the measured partition coefficients using solute retention times and the cell utilized partitioning model to account for matrix effects in the following optimization experiments. Furthermore, the carbon–carbon linked oligomers in the lignin oil substrates elute together and separately from the aromatic monomers in lignin oil, resulting in an oligomer-rich product stream. Case studies of optimization of poplar RCF-derived lignin oil separations exhibited non-polar monomer yields of 95–99% with purities of 72–96%. Additionally, the same separation using a propyl-rich lignin oil produced from a H2-free RCF process showed a nearly 46% increase in normalized productivity, exhibiting the importance of tuning feed composition to improve separation performance. Taken together, this work shows that CCC is a promising method for simultaneous lignin monomer–monomer and monomer–oligomer separations. [ABSTRACT FROM AUTHOR]

Essential oils (EOs) are vital secondary metabolites in plants. They have garnered substantial attention owing to their distinct flavors and desirable attributes, including potent antioxidant, antibacterial, and antitumor properties. Nevertheless, the active constituents of EOs exhibit intricate chemical structures, and conventional separation techniques are inadequate for purifying the individual chemical components from EOs. High-speed countercurrent chromatography, based on the principles of a hydrodynamic equilibrium system, has emerged as a liquid–liquid chromatographic separation method renowned for its ability to handle substantial single injection volumes and the absence of irreversible adsorption. Consequently, in recent years, this technique has been widely employed in the isolation and refinement of natural products. In this review, a comprehensive analysis is conducted, contrasting the merits and demerits of high-speed countercurrent chromatography with conventional separation methods. The solvent systems, elution modes, commonly employed detectors, and practical applications are reviewed in the context of high-speed countercurrent chromatography for essential oil separation and purification. Furthermore, this review offers a glimpse into the potential prospects of applying this technique, with the intention of serving as a valuable reference for the use of high-speed countercurrent chromatography in the purification of EOs. [ABSTRACT FROM AUTHOR]

Crocin-I, a valuable natural compound found in saffron (Crocus sativus L.), is the most abundant among the various crocin structures. Developing a cost-effective and scalable purification process to produce high-purity crocin-I is of great interest for future investigations into its biological properties and its potential applications in the treatment of neurological disorders. However purifying crocin-I through single-column preparative chromatography (batch) poses a yield-purity trade-off due to structural similarities among crocins, meaning that the choice of the collection window sacrifices either yield in benefit of higher purity or vice versa. This study demonstrates how the continuous countercurrent operating mode resolves this dilemma. Herein, a twin-column MCSGP (multicolumn countercurrent solvent gradient purification) process was employed to purify crocin-I. This study involved an environmentally friendly ethanolic extraction of saffron stigma, followed by an investigation into the stability of the crocin-I within the feed under varying storage conditions to ensure a stable feed composition during the purification. Then, the batch purification process was initially designed, optimized, and subsequently followed by the scale-up to the MCSGP process. To ensure a fair comparison, both processes were evaluated under similar conditions (e.g., similar total column volume). The results showed that, at a purity grade of 99.7%, the MCSGP technique demonstrated significant results, namely + 334% increase in recovery + 307% increase in productivity, and − 92% reduction in solvent consumption. To make the purification process even greener, the only organic solvent employed was ethanol, without the addition of any additive. In conclusion, this study presents the MCSGP as a reliable, simple, and economical technique for purifying crocin-I from saffron extract, demonstrating for the first time that it can be effectively applied as a powerful approach for process intensification in the purification of natural products from complex matrices. [ABSTRACT FROM AUTHOR]

The primary goals of green analytical chemistry (GAC) involve the development of novel analytical technologies or the modification of existing methods to incorporate procedures with reduced reliance on hazardous chemicals. Several approaches can be employed to achieve these objectives, including the utilization of environmentally friendly solvents and reagents, shortening chromatographic separation times, and the miniaturization of analytical devices. The review summarizes the evolution of GAC with its specific principles and with special emphasis on recent applications in modern era. A detailed exploration of the choice of green solvents and their practical utilization in green liquid chromatography (LC) techniques for environmentally friendly analysis of pharmaceuticals has been discussed. An ordinary description with a tabular illustration of various modern green analytical techniques like micellar LC, superheated water chromatography, high‐speed LC, enhanced fluidity LC, two‐dimensional LC, and aspects of gas chromatography has been thoroughly provided for concepts with their mechanism and limitations. GAC provides potential applications for food analysis, environmental analysis of pharmaceutically active compounds in the aquatic environment with special emphasis on strategies for greening LC and impurity profiling, and so on. Numerous benefits, limitations, and challenges that are linked to the field of GAC are highlighted in this review. We pay special attention to new green approaches, such as automation, miniaturization, direct analysis techniques, and some initiatives like the National Environmental Methods Index Label for Analytical Eco‐Scale assessment. In spite of many promising advancements, the understanding is still limited and challenging. [ABSTRACT FROM AUTHOR]

An effective method by high‐speed countercurrent chromatography coordinated with silver nitrate for the preparative separation of sterones and triterpenoid saponins from Achyranthes bidentata Blume was developed. Methyl tert‐butyl ether/n‐butanol/acetonitrile/water (4:2:3:8, v/v/v/v) was selected for 20‐hydroxyecdysone (compound 1), chikusetsusaponin IVa methyl ester (compound 4), 2′‐glycan‐11‐keto‐pigmented saponin V (compound 5), as well as a pair of isomers of 25S‐inokosterone (compound 2) and 25R‐inokosterone (compound 3), which were further purified by silver nitrate coordinated high‐speed countercurrent chromatography. What is more, dichloromethane/methanol/isopropanol/water (6:6:1:4, v/v/v/v) was applied for calenduloside E (compound 6), 3β‐[(O‐β‐d‐glucuronopyranosyl)‐oxy]‐oleana‐11,13‐dien‐28‐oic acid (compound 7), zingibroside R1 (compound 8) and chikusetsusaponin IVa (compound 9). Adding Ag+ to the solvent system resulted in unique selectivity for 25R/25S isomers of inokosterone, which increased the complexing capability and stability of Ag+ coordinated 25S‐inokosterone, as well as the α value between them. These results were further confirmed by the computational calculation of geometry optimization and frontier molecular orbitals assay. Comprehensive mass spectrometry and nuclear magnetic resonance analysis demonstrated the structures of the obtained compounds. [ABSTRACT FROM AUTHOR]

In recent years, discovering new drug candidates has become a top priority in research. Natural products have proven to be a promising source for such discoveries as many researchers have successfully isolated bioactive compounds with various activities that show potential as drug candidates. Among these compounds, phenolic compounds have been frequently isolated due to their many biological activities, including their role as antioxidants, making them candidates for treating diseases related to oxidative stress. The isolation method is essential, and researchers have sought to find effective procedures that maximize the purity and yield of bioactive compounds. This review aims to provide information on the isolation or separation methods for phenolic compounds with antioxidant activities using column chromatography, medium-pressure liquid chromatography, high-performance liquid chromatography, counter-current chromatography, hydrophilic interaction chromatography, supercritical fluid chromatography, molecularly imprinted technologies, and high-performance thin layer chromatography. For isolation or purification, the molecularly imprinted technologies represent a more accessible and more efficient procedure because they can be applied directly to the extract to reduce the complicated isolation process. However, it still requires further development and refinement. [ABSTRACT FROM AUTHOR]

Spondias dulcis Parkinson have been used in traditional medicine in Asia, Oceania, and South America, for different diseases conditions and as a functional food. The scientific literature described as different potential pharmacology such as antioxidant, anti-inflammatory, antimicrobial, thrombolytic and enzymatic inhibitor. This study aimed to: (1) establish the pharmacological activity in intestinal motility in vivo and antioxidant activity in vitro; (2) perform the acute toxicology test in mouse; (3) characterize the phytochemical profile based on counter-current chromatography (CCC) and NMR analysis. The results revealed a laxative effect of S. dulcis extract and a high antioxidant activity (IC50 = 5.10 for DPPH assay and 14.14 for hydrogen peroxide scavenging test). No side effects were observed in the oral acute toxicity test for a dose up to 2000 mg/kg. The chemical profile was identified by CCC and NMR, and the comparison of the data obtained with previous literature revealed the presence of the flavonoid rutin (Quercetin-3-O-rutinoside) in the extract. [ABSTRACT FROM AUTHOR]

Accurate screening and targeted preparative isolation of active substances in natural medicines have long been two technical challenges in natural medicine research. This study outlines a new approach to improve the efficiency of natural product preparation, focusing on rapidly and accurately screening potential active ingredients in Inonotus obliquus as well as efficiently preparing 5‐lipoxidase (5‐LOX) inhibitors, to provide new ideas for the treatment of asthma with Inonotus obliquus. First, we used ultrafiltration (UF) mass spectrometry to screen for three potential inhibitors of 5‐LOX in Inonotus obliquus. Subsequently, the inhibitory effect of the active ingredients screened in the UF assay on 5‐LOX was verified using the molecular docking technique, and the potential role of the active compounds in Inonotus obliquus for the treatment of asthma was analyzed by network pharmacology. Finally, based on the above activity screening guidelines, we used semi‐preparative liquid chromatography and consecutive high‐speed countercurrent chromatography to isolate three high‐purity 5‐LOX inhibitors such as betulin, lanosterol, and quercetin. Obviously, through the above approach, we have seamlessly combined rapid discovery, screening, and centralized preparation of the active ingredient with molecular‐level interactions between the active ingredient and the protease. [ABSTRACT FROM AUTHOR]

Continuous chromatography has emerged as one of the most attractive methods for protein purification. Establishing such systems involves installing several chromatographic units in series to enable continuous separation processes and reduce the cost of the production of expensive proteins and biopharmaceuticals (such as monoclonal antibodies). However, most of the established systems are bulky and plagued by high dead volume, which requires further optimization for improved separation procedures. In this article, we present a miniaturized periodic counter-current chromatography (PCCC) system, which is characterized by substantially reduced dead volume when compared to traditional chromatography setups. The PCCC device was fabricated by 3D printing, allowing for flexible design adjustments and rapid prototyping, and has great potential to be used for the screening of optimized chromatography conditions and protocols. The functionality of the 3D-printed device was demonstrated with respect to the capture and polishing steps during a monoclonal antibody purification process. Furthermore, this novel miniaturized system was successfully used for two different chromatography techniques (affinity and ion-exchange chromatography) and two different types of chromatographic units (columns and membrane adsorbers). This demonstrated versability underscores the flexibility of this kind of system and its potential for utilization in various chromatography applications, such as direct product capture from perfusion cell cultures. [ABSTRACT FROM AUTHOR]

Tar sand, an aggregate of sand, clay, water and bitumen was sampled from Okitipupa area of Ondo State, Nigeria, extracted and optimized using n-hexane and toluene in a soxhlet extractor todetermine solvent effectiveness in extracting the bitumen content and their environmental effect. The experiment was carried out by heating the solvent at and above their boiling temperatures solely to get an optimum yield at a temperature and time of extraction of oil from the tar sand. The analysis gave an optimal yield of 9.07 % at 75oC in 4 h when n-hexane was used and an optimal yield of 21.27 % at 120oC in 4 h when toluene was used. A drastic change in colour was also notice in the sand residue to almost sandy soil nature when toluene was used as solvent compare to when n-hexane was used which shows a slight change from its blackish nature. The change in colour of the black tar sand is an indication of high extraction efficiency. Hence, owing to high demand for conventional oil supply in the country and the urgent need for an alternative source of oil, the bitumen extract can serve as an alternative source of unconventionaloil supply in Nigeria and feedstock in refineries. [ABSTRACT FROM AUTHOR]

AbstractContext The antidiabetic effects of flavonoids have been reported, but it is still unclear whether 5,7,3′,4′,5′-pentamethoxyflavone, isolated from Bauhinia championii Benth. (Fabaceae), also exhibits such properties.Objective To isolate 5,7,3′,4′,5′-pentamethoxyflavone from B. championii using high-speed countercurrent chromatography and examine its potential in treating diabetic nephropathy.Materials and methods The phytochemical constituents from the stems of B. championii were separated and purified with high-speed countercurrent chromatography; 5,7,3′,4′,5′-pentamethoxyflavone (PMF) was identified by mass spectrum, 1H-NMR, and 13C-NMR. After exposing mesangial cells to 30 mM glucose and either 5 μM or 10 μM PMF for 6 h, the levels of fibronectin (FN) and p-Smad2/3 were analyzed using Western blotting. Male Sprague–Dawley rats were injected intraperitoneally with 55 mg/kg streptozotocin to induce diabetes and then were randomized into three groups (n = 10): vehicle administration, low-dose (5 mg/kg) PMF, and high-dose (25 mg/kg) PMF by intragastric gavage for 3 months. A healthy group was included as the control.Results Compared to the diabetic group, low-dose and high-dose PMF treatment decreased the phosphorylation of Smad2/3 by 0.54- and 0.52-fold, and the accumulation of FN decreased by 0.82- and 0.77-fold in vitro; the phosphorylation of Smad2/3 was decreased by 0.39- and 0.37-fold, and the accumulation of FN decreased by 0.47- and 0.40-fold in vivo, respectively. Furthermore, PMF alleviated the glomerular basement membrane thickness and foot process fusion.Conclusion The findings suggest for the first time that PMF may be a promising treatment option for diabetic kidney fibrosis, which warrants additional clinical investigation.

German chamomile is one of the most effective herbal elements used in anti‐allergic products and as an antioxidant. Herein, the antioxidant activity of different extract fractions of German chamomile was initially evaluated using an off‐line 2,2‐diphenyl‐1‐picrylhydrazyl spectrophotometric assay. The ethyl acetate extract demonstrated the highest efficacy in scavenging free radicals. Based on this, a rapid screening and separation method using ultra‐high‐performance liquid chromatography combined with the 2,2‐diphenyl‐1‐picrylhydrazyl assay was implemented to identify antioxidants in the ethyl acetate fraction of German chamomile flowers. Ten potential radical scavengers were tentatively screened from German chamomile using a target‐guided isolating approach with off‐line two‐dimensional high‐speed countercurrent chromatography and the structures of the compounds were analyzed and identified. Ultimately, 10 radical scavengers were obtained from the ethyl acetate extract with a purity quotient exceeding 90%. The results demonstrated the effectiveness and reproducibility of this method for isolating potential antioxidants from complex mixtures in a targeted manner. This strategy can be applied to the target‐guided isolation of complex mixtures of natural products with broad K‐values and similar structures. [ABSTRACT FROM AUTHOR]

In the present study, twelve compounds from Dioscorea spongiosa were successfully purified by an efficient technique combined bioassay‐guided fractionation macroporous resin column chromatography (MRCC) pretreatment and high‐speed counter‐current chromatography (HSCCC) separation for the first time. Then, D101 MRCC was used to fractionate the crude extract into five parts, which further applied the bioassay‐guided fractionation strategy to screen the active fractions of 2 and 4. As for the separation, 200 mg Fr.2 was purified by HSCCC using EtOAc/n‐BuOH/H2O (2:2:3, v/v), leading to annulatomarin (1), dioscoresides C (2), diosniponol C (3), methyl protodioscin (4), pseudoprotodioscin (5), protogracillin (6), as well as 200 mg Fr.4 yielding montroumarin (7), dioscorone A (8), diosniponol D (9), protodioscin (10), gracillin (11), and dioscin (12) using CH2Cl2/MeOH/H2O (3:3:2, v/v) with the purities over 95.0%. Finally, the isolates were assayed for their anti‐inflammatory, urico‐lowering, and anti‐diabetic activities in vitro, which indicated that the steroidal saponins of 5, 6, and 11 showed all these three activities. [ABSTRACT FROM AUTHOR]

Countercurrent chromatography (CCC) is a potent separation approach known for its remarkable efficiency and capacity in preparation. It's applied as a substitute or combined with different chromatographic techniques, resulting in its rebranding as multidimensional CCC (MDCCC). Numerous essential mixtures from natural products contain hundreds or thousands of distinct components of importance. These mix types are too complicated to separate in any reasonable time using a single CCC dimension. However, if a multidimensional technique is utilized, where a complex mixture is separated by an initial dimension, smaller fractions of that separation are gathered. Each fraction is studied individually; complex mixes can be resolved relatively quickly. Thus, several MDCCC separation features have been studied to demonstrate their advantages, limitations, and prospective capacity to separate exceedingly complex mixtures. In this review, MDCCC aspects, including principles, multiple columns system, multilayer coil J‐type, on‐line monitoring system, and applications, have been thoroughly_explored. [ABSTRACT FROM AUTHOR]

Myrica rubra (Lour.) Siebold & Zucc bark is a traditional natural medicine used by the people of the Dong minority in western Hunan in China. In this study, the main compounds in Myrica rubra bark including epigallocatechin gallate, myricetrin, myricetin, taraxerol, myricanol, and 11-O-acetylmyricanol were separated using both silica gel column chromatography and high-speed countercurrent chromatography (HSCCC). Notably, it is the first report of discovering 11-O-acetylmyricanol from Myrica rubra bark. The results of the bioactivity studies suggested that epigallocatechin gallate showed the highest α-glucosidase inhibitory activity, while myricetin exhibited the highest reactive oxygen species (ROS) scavenging ability in zebrafish embryos. Intriguingly, myricanol exhibited strong apoptosis-inducing activity on HepG2 cells, and further studies revealed that myricanol was capable of promoting the cleavage of caspase 3, 8, and 9, then resulting in the apoptosis in HepG2 cells. The findings of the present study have important implications for the separation of the main compounds in Myrica rubra and will provide credence to the ethnomedicinal application of the isolated compounds against cardiovascular disease and cancer. [ABSTRACT FROM AUTHOR]

Endemic in 21 countries, Chagas disease, also known as American Trypanosomiasis, is a neglected tropical disease (NTD) caused by the protozoan parasite Trypanosoma cruzi. The available drugs for the treatment of this disease, benznidazole and nifurtimox, are outdated and display severe side effects. Thus, the discovery of new drugs is crucial. Based on our continuous studies aiming towards the discovery of natural products with anti-T. cruzi potential, the MeOH extract from aerial parts of Baccharis sphenophylla Dusén ex. Malme (Asteraceae) displayed activity against this parasite and was subjected to high-performance countercurrent chromatography (HPCCC), to obtain one unreported syn-labdane diterpene — sphenophyllol (1) — as well as the known compounds gaudichaudol C (2), ent-kaurenoic acid (3), hispidulin (4), eupafolin (5), and one mixture of di-O-caffeoylquinic acids (6–8). Compounds 1–8 were characterized by analysis of nuclear magnetic resonance (NMR) and mass spectrometry (MS) data. When tested against trypomastigote forms, isolated labdane diterpenes 1 and 2 displayed potent activity, with EC50 values of 20.1 μM and 2.9 μM, respectively. The mixture of chlorogenic acids 6–8, as well as the isolated flavones 4 and 5, showed significant activity against the clinically relevant amastigotes, with EC50 values of 24.9, 12.8, and 2.7 μM, respectively. Nonetheless, tested compounds 1–8 displayed no cytotoxicity against mammalian cells (CC50 > 200 μM). These results demonstrate the application of HPCCC as an important tool to isolate bioactive compounds from natural sources, including the antitrypanosomal extract from B. sphenophylla, allowing for the development of novel strategic molecular prototypes against tropical neglected diseases. [ABSTRACT FROM AUTHOR]

Centrifugal partition chromatography in the pH‐zone‐refining mode was successfully applied to the separation of alkaloids from the crude extract of Corydalis decumbens. The experiment was performed with a two‐phase solvent system composed of petroleum ether‐ethyl acetate‐ethanol‐water (5:5:3:7, v/v/v/v) where triethylamine (10 mM) was added to the stationary phase and hydrochloric acid (10 mM) to the mobile phase. From 1.6 g of the crude extract, 43 mg protopine, 189 mg (+)‐egenine, and 158 mg tetrahydropalmatine were obtained with a purity of 98.2%, 94.6%, and 96.7%, respectively. Tetrahydropalmatine showed an interesting anticomplement effect with CH50 0.11 and AP50 0.25 mg/mL, respectively. In a mechanistic study, tetrahydropalmatine interacted with C1, C3, C4, and C5 components in the complement activation cascade. [ABSTRACT FROM AUTHOR]

Meconopsis integrifolia (Maxim.) Franch. is used extensively in traditional Tibetan medicine for its potent anti‐inflammatory properties. In this study, six cyclooxygenase‐2 (COX‐2) inhibitors were purified from M. integrifolia using high‐speed counter‐current chromatography guided by ultrafiltration liquid chromatography (ultrafiltration‐LC). First, ultrafiltration‐LC was performed to profile the COX‐2 inhibitors in M. integrifolia. The reflux extraction conditions were further optimized using response surface methodology, and the results showed that the targeted COX‐2 inhibitors could be well enriched under the optimized extraction conditions. Then the six target COX‐2 inhibitors were separated by high‐speed countercurrent chromatography with a solvent system composed of ethyl acetate/n‐butanol/water (4:1:4, v/v/v. Finally, the six COX‐2 inhibitors, including 21.2 mg of 8‐hydroxyluteolin 7‐sophoroside, 29.6 mg of 8‐hydroxyluteolin 7‐[6′''‐acetylallosyl‐(1→2)‐glucoside], 42.5 mg of Sinocrassoside D3, 54.1 mg of Hypolaetin 7‐[6′′′‐acetylallosyll‐(l→2)‐3′′‐acetylglucoside, 30.6 mg of Hypolaetin 7‐[6′′′‐acetylallosyll‐(l→2)‐6′′‐acetylglucoside and 17.8 mg of Hypolaetin were obtained from 500 mg of sample. Their structures were elucidated by 1H‐NMR spectroscopy. This study reveals that ultrafiltration‐LC combined with high‐speed counter‐current chromatography is a robust and efficient strategy for target‐guided isolation and purification of bioactive molecules. It also enhances the scientific understanding of the anti‐inflammatory properties of M. integrifolia but also paves the way for its further medicinal applications. [ABSTRACT FROM AUTHOR]

Poria Cum Radix Pini is a rare medicinal fungus that contains several potential therapeutic ingredients. On this basis, a particle swarm mathematical model was used to optimize the extraction process of total triterpenes from P. Cum Radix Pini, and xanthine oxidase inhibitors were screened using affinity ultrafiltration mass spectrometry. Meanwhile, the accuracy of the ultrafiltration assay was verified by molecular docking experiments and molecular dynamics analysis, and the mechanism of action of the active compounds for the treatment of gout was analyzed by enzymatic reaction kinetics and network pharmacology. A high‐speed countercurrent chromatography method combined with the consecutive injection and the economical two‐phase solvent system preparation using functional activity coefficient of universal quasichemical model (UNIFAC) mathematical model was developed for increasing the yield of target compound. In addition, dehydropachymic acid and pachymic acid were used as competitive inhibitors, and 3‐O‐acetyl‐16alpha‐hydroxydehydrotrametenolic acid and dehydrotrametenolic acid were used as mixed inhibitors. Then, activity‐oriented separation and purification were performed by high‐speed countercurrent chromatography combined with semi‐preparative high‐performance liquid chromatography and the purity of the four compounds isolated was higher than 90%. It will help to provide more opportunities to discover and develop new potential therapeutic remedies from health care food resources. [ABSTRACT FROM AUTHOR]

α-Amylase inhibitors constitute a category of compounds capable of diminishing α-amylase activity and hold significant promise for the development of novel drugs. This study focused on assessing the potential of Schisandra chinensis Baill extract to inhibit α-amylase. However, sieving and segregating inhibitors from intricate mixtures presents an obstacle. Consequently, this study devised and refined a methodology centered on immobilized α-amylase to identify and isolate natural inhibitors from S. chinensis. The immobilized α-amylase, featuring a load capacity of 254.18 ± 1.20 μg/mg and a specific activity of 2.69 ± 0.01 U/mg, was achieved through meticulous optimization of preparation conditions. Subsequently, this immobilized enzyme was used to isolate bioactive compounds from S. chinensis extract. The results revealed the presence of an active compound with an IC 50 of 324.12 ± 11.23 μg/mL. This compound was isolated via magnetic immobilized enzyme-based ligand fishing, employing high-speed countercurrent chromatography, and was conclusively identified as 5-hydroxymethylfurfural. The molecular docking analyses revealed its ability to interact with the active pocket of α-amylase through multiple hydrogen bonds. [Display omitted] • α-Amylase was immobilized on mesoporous silica particles for ligand fishing. • Main active component was screened from S. chinensis by immobilized α-amylase. • 5-Hydroxymenthyfurfural was the main active component in S. chinensis fruit. • The interaction between active compound and α-amylase was investigated. [ABSTRACT FROM AUTHOR]

Introduction: Many secondary metabolites isolated from plants have been described in the literature owing to their important biological properties and possible pharmacological applications. However, the identification of compounds present in complex plant extracts has remained a great scientific challenge, is often laborious, and requires a long research time with high financial cost. Objectives: The aim of this study was to develop a method that allows the identification of secondary metabolites in plant extracts with a high degree of confidence in a short period of time. Material and methods: In this study, an ethanolic extract of Coffea arabica leaves was used to validate the proposed method. Countercurrent chromatography was chosen as the initial step for extraction fractionation using gradient elution. Resulting fractions presented a variation of compounds concentrations, allowing for statistical total correlation spectroscopy (STOCSY) calculations between liquid chromatography coupled with high‐resolution tandem mass spectrometry (LC‐HRMS/MS) and NMR across fractions. Results: The proposed method allowed the identification of 57 compounds. Of the annotated compounds, 20 were previously described in the literature for the species and 37 were reported for the first time. Among the inedited compounds, we identified flavonoids, alkaloids, phenolic acids, coumarins, and terpenes. Conclusion: The proposed method presents itself as a valid alternative for the study of complex extracts in an effective, fast, and reliable way that can be reproduced in the study of other extracts. A novel method was developed to identify secondary metabolites in plant extracts with high confidence and efficiency. Using an ethanolic extract of Coffea arabica leaves, countercurrent chromatography was employed for extraction fractionation, followed by statistical total correlation spectroscopy (STOCSY) calculations between liquid chromatography coupled with high‐resolution tandem mass spectrometry (LC‐HRMS/MS) and NMR. This approach successfully identified 57 compounds, with 20 known and 37 newly reported compounds, including flavonoids, alkaloids, phenolic acids, coumarins, and terpenes. [ABSTRACT FROM AUTHOR]

Hydrophilic deep eutectic solvents are actively positioned as efficient extractants for removing heterocyclic compounds from light hydrocarbon fractions. Of particular interest is the subclass of natural deep eutectic solvents (NaDESs), since they contain substances of exclusively natural origin. However, these processes have not been systematically studied to date in extraction equipment. To study the process of countercurrent extraction of pyridine, quinoline, and indole from a model solution of light hydrocarbon fractions using commercial equipment, a series of NaDESs based on citric and malic acids, xylitol, and water was used for the first time in this work. The high extraction capacity of these NaDES was demonstrated in laboratory experiments, and the extraction mechanism was determined. A detailed study of the efficiency of extraction of heterocycles with varying process conditions allowed us to move on to studying the process using extractors of the mixer–settler type. From the model solution of light hydrocarbon fractions, pyridine, quinoline, and indole were removed to concentrations <1 ppm by countercurrent extraction using a cascade of six mixer–settlers. [ABSTRACT FROM AUTHOR]

This article proposes a solvent system screening strategy for compounds with similar UV absorption in complex samples by UV spectrophotometer. There is no need to calculate the partition coefficient value of each compound, only the partition coefficient of the whole sample. The partition coefficient value should be close to 1 in order to obtain as many high‐speed counter‐current chromatography fractions as possible. Then, preparative HPLC was used to purify the high‐speed counter‐current chromatography fractions. Based on the above strategy, seven c‐glycosyl flavonoids and an amino acid were successfully obtained from barley seedlings through high‐speed counter‐current chromatography fractionation with ethyl acetate/n‐butanol/water (8:2:10, v:v:v) system followed by preparative HPLC purification. The research shows that high‐speed counter‐current chromatography could be well developed as a tool for fractionation before purification, and greatly improves the separation efficiency. [ABSTRACT FROM AUTHOR]

An instrumental neutron activation analysis method in conjunction with Compton suppression spectrometry was developed for the simultaneous determination of nanogram levels of Cl, Br and I in salmon muscle and skin. Several non-chlorinated solvent systems were evaluated for the extraction of lipids containing organohalogens and an acetone-hexane (1:1) system was selected because it gave the highest extraction efficiency with least contamination. An extensive quality assurance program was designed. Lipid fractionation procedures were developed using size exclusion, solid phase extraction and thin layer chromatography followed by characterization using 1H, 13C and 31P nuclear magnetic resonance and mass spectrometry. Results show that both muscle and skin had high levels of triacylglycerols. [ABSTRACT FROM AUTHOR]

The antidiabetic effects of flavonoids have been reported, but it is still unclear whether 5,7,3′,4′,5′-pentamethoxyflavone, isolated from Bauhinia championii Benth. (Fabaceae), also exhibits such properties. To isolate 5,7,3′,4′,5′-pentamethoxyflavone from B. championii using high-speed countercurrent chromatography and examine its potential in treating diabetic nephropathy. The phytochemical constituents from the stems of B. championii were separated and purified with high-speed countercurrent chromatography; 5,7,3′,4′,5′-pentamethoxyflavone (PMF) was identified by mass spectrum, 1H-NMR, and 13C-NMR. After exposing mesangial cells to 30 mM glucose and either 5 μM or 10 μM PMF for 6 h, the levels of fibronectin (FN) and p-Smad2/3 were analyzed using Western blotting. Male Sprague–Dawley rats were injected intraperitoneally with 55 mg/kg streptozotocin to induce diabetes and then were randomized into three groups (n = 10): vehicle administration, low-dose (5 mg/kg) PMF, and high-dose (25 mg/kg) PMF by intragastric gavage for 3 months. A healthy group was included as the control. Compared to the diabetic group, low-dose and high-dose PMF treatment decreased the phosphorylation of Smad2/3 by 0.54- and 0.52-fold, and the accumulation of FN decreased by 0.82- and 0.77-fold in vitro; the phosphorylation of Smad2/3 was decreased by 0.39- and 0.37-fold, and the accumulation of FN decreased by 0.47- and 0.40-fold in vivo, respectively. Furthermore, PMF alleviated the glomerular basement membrane thickness and foot process fusion. The findings suggest for the first time that PMF may be a promising treatment option for diabetic kidney fibrosis, which warrants additional clinical investigation. [ABSTRACT FROM AUTHOR]

Polymethoxyflavones were a unique class of natural and safe flavonoids containing two or more methoxy groups, which were also the most abundant edible part in Citrus peel. The optimum condition in the process of selective extraction of polymethoxylated flavones from Citrus peel by matrix solid‐phase dispersion (MSPD) was as follows: SBA‐15 as adsorbent, ethyl acetate as eluent, the mass ratio of adsorbent to sample 1:1, and the mixture of sample and adsorbent was ground for 3 min. Twelve antioxidants were successfully screened by micro‐fractionation bioactivity evaluation assay, in which four of them were flavonoid glycosides, seven of them were polymethoxylated flavones, and one was phenylpropanoid. 1‐sinapoly‐β‐D‐glucopyranoside (1) was reported for the first time in Citrus peel. And antioxidant capacity of 1‐sinapoly‐β‐D‐glucopyranoside, 5, 7, 8, 3′, 4′, 5′‐hexamethoxyflavone (6), hexamethoxyflavone (11), and 5, 6, 7, 4′‐tetramethoxyflavone (7) were reported for the first time. Nobiletin (compound 8), 3, 5, 6, 7, 8, 3′, 4′‐heptamethoxyflavone (9) and tangeretin (10) were isolated and purified by countercurrent chromatography combined with preparative liquid chromatography. Antioxidant activity evaluation indicated that the three isolated polymethoxylated flavones owned similar antioxidant activity. This study indicated that MSPD combined with micro‐fractionation bioactive evaluation was efficient in screening bioactive compounds for rapid extraction and effective pinpointing bioactive substances in natural products. [ABSTRACT FROM AUTHOR]

In this work, eight compounds from Phellodendron chinense were separated and purified by pH‐zone refining counter‐current chromatography and traditional counter‐current chromatography coupled with online‐storage inner‐recycling counter‐current chromatography (IRCCC). The pH‐zone‐refining mode was adopted for separating 2.0 g of crude extract with the solvent system of chloroform–methanol–water (4:3:3, v/v), in which 10 mM hydrochloric acid and 10 mM triethylamine were added in the stationary and mobile phases, respectively. Meanwhile, traditional counter‐current chromatography coupled with online‐storage IRCCC separation was performed by the solvent system of n‐hexane‐ethyl acetate‐methanol‐water (5:5:2:8, v/v). Finally, eight compounds, including six alkaloids as 6‐methylpiperidin‐2‐one(1), isoplatydesmine(4), berlambine(5), epiberberine(6), palmatine(7), berberine(8) and two phenolic acids as ferulic acid(2), isoferulic acid(3), were successfully obtained using these three different CCC modes with the purities over 95.0%. [ABSTRACT FROM AUTHOR]

This research investigated the effectiveness of an integrated method for the extraction and separation of naphthoquinones and diarylheptanes from exocarp of Juglands mandshurica Maxim. (namely, green walnut husks). The target compounds were obtained by ultra‐turrax homogenization (UTH) coupled with ultrasound‐assisted extraction (UAE) technology followed by high‐speed countercurrent chromatography (HSCCC). The UTH‐UAE extraction method achieved higher efficiency with 2.49‐ and 2.36‐fold to those by UAE, and 1.39‐ and 1.34‐fold to those by UTH in a short time. HSCCC was adopted for further separation and purification; six target compounds, namely, regiolone (RE), juglone (JU), myricatomento‐genin (MG), galleon (GA), 2‐oxatrycyclo[13.2.2.13,7]eicosa‐3,5,7(20),15,17,18‐hexaen‐10‐16‐diol (OE), and juglanin A (JA), were separated with more than 95.37% purities and more than 84.71% final recovery rates, respectively. In this study, the integrated strategy of extraction and separation could get high purity compounds quickly, which would provide time and solvent saved method for the natural products separation from plants. [ABSTRACT FROM AUTHOR]

Grapevine canes are an important source of bioactive compounds, such as stilbenoids. This study aimed to evaluate an in silico method, based on the Conductor-like Screening Model for Real Solvents (COSMO-RS) to isolate stilbenoids from a grapevine cane extract by offline heart-cut high-performance countercurrent chromatography (HPCCC). For the following extraction of resveratrol and ε-viniferin from grapevine canes, natural deep eutectic solvents (NADES) were used as an environmentally friendly alternative to the traditionally used organic solvents. In order to evaluate a variety of combinations of hydrogen bond acceptors (HBAs) and hydrogen bond donors (HBDs) for the targeted extraction of stilbenoids, COSMO-RS was applied. In particular, ultrasonic-assisted extraction using a solvent mixture of choline chloride/1,2-propanediol leads to higher extraction yields of resveratrol and ε-viniferin. COSMO-RS calculations for NADES extraction combined with HPCCC biphasic solvent system calculations are a powerful combination for the sustainable extraction, recovery, and isolation of natural products. This in silico-supported workflow enables the reduction of preliminary experimental tests required for the extraction and isolation of natural compounds. [ABSTRACT FROM AUTHOR]