Macroporous Resin Research Articles

Screening of anti-cancer compounds from Vaccariae Semen by lung cancer A549 cell fishing and UHPLC-LTQ Orbitrap MS

Vaccariae Semen, derived from the dried ripe seed of Vaccaria segetalis (Neck.) Garcke, has various therapeutic characteristics in traditional Chinese medicine (TCM), containing promoting blood circulation and unblocking meridians. It exhibits significant anti-cancer activity and is therapeutically utilized to treat and reduce chemotherapy adverse effects in cancer patients, notably those with lung cancer. However, the active ingredients responsible for its anti-lung cancer efficacy remain unknown. In this study, we used A549 cell fishing in conjunction with UHPLC-LTQ Orbitrap MS to screen for anti-lung cancer active components in Vaccariae Semen. The cell counting Kit-8 (CCK-8) assay revealed that the n-butanol extract substantially reduced A549 cell growth. Through the cell fishing assay, we found 14 A549 cell-binding compounds in the n-butanol extract, all of which were identified as triterpenoid saponins. The total saponins of Vaccariae Semen were subsequently purified using macroporous adsorption resin (MAR), and they showed a significant inhibitory effect on the proliferation of A549 lung cancer cells, as well as alterations in cell morphology, apoptosis, and fragmentation. In conclusion, saponins were discovered as the key active components responsible for the anti-lung cancer activity of Vaccariae Semen.

Adsorptive recovery of polyphenols from apple pomace

This study was carried out to investigate the influence of different operating parameters such as pH, amount of adsorbent, temperature, adsorbent-to-apple pomace (AP) ratio, and type of elution solvent on the recovery of polyphenols from AP. The study used a non-functionalized macroporous polymetric food-grade resin (Amberlite FPX66) as an adsorbent for the recovery of polyphenols from apple pomace (AP) on a laboratory scale. Adsorption was carried out using a continuous process and desorption was carried out using a batch process. The adsorption on the resin was influenced by the pH and the AP: resin ratio. At pH 4, for the first 30 min, about 94% of polyphenols were adsorbed onto the 5 g resin, and about 77 and 82% of the compounds were adsorbed onto the resin at pH 2 and pH 7, respectively. After 150 min, about 86% of polyphenols were adsorbed onto the 5 g resin, whereas the polyphenols adsorbed on the 10 g and 20 g were 71 and 64%, respectively. However, for the first 30 min, 10 g and 20 g polyphenols were fully adsorbed onto the resin. Desorption studies were carried out using a resin that was taken from the batch process. Elution for the desorption studies was carried out using different solvents (water, ethanol, and isopropanol) at different temperatures ranging from 40 to 180 °C to determine their influence on the recovery of individual polyphenols. Isopropanol showed more recovery for polyphenols at 40 °C compared to when water and methanol were used. The study shows that Amberlite FPX 66 has the capability to recover polyphenol, however, the operational temperature should be considered.

Heterocyclic compounds and phenolic glycosides from flowers of Dendrobium officinale

Eight heterocyclic compounds and twelve phenolic glycosides were separated from the water extract of Dendrobium officinale flowers through chromatographic techniques, such as Diaion HP-20 macroporous adsorption resin column chromatography(CC), silica gel CC, ODS CC, Sephadex LH-20 CC, and preparative high performance liquid chromatography(PHPLC). According to the spectroscopic analyses(MS, ~1H-NMR, and ~(13)C-NMR) and optical rotation data, the compounds were identified as dendrofurfural A(1), 2'-deoxyadenosine(2), 4-[2-formyl-5-(hydroxymethyl)-1H-pyrrol-1-yl] butanoic acid(3), 4-[2-formyl-5-(methoxymethyl)-1H-pyrrol-1-yl] butanoic acid(4), 1-(2-hydroxyethyl)-5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde(5), 5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde(6), methyl 5-(hydroxymethyl)-furan-2-carboxylate(7),(S)-5-hydroxymethyl-5H-furan-2-one(8), 2-methoxyphenyl-1-O-β-D-glucopyranoside(9), arbutin(10), isotachioside(11), 2,6-dimethoxy-4-hydroxyphenol-1-O-β-D-glucopyranoside(12), orcinol glucoside(13), tachioside(14), gastrodin(15), 4-O-β-D-glucopyranosylvanillyl alcohol(16), 2,6-dimethoxy-4-hydroxymethylphenol-1-O-β-D-glucopyranoside(17), icariside D_2(18), 4-formylphenyl-β-D-glucopyranoside(19), and vanillin-4-O-β-D-glucopyranoside(20). Among them, compound 1 is a new furfural benzyl alcohol condensate, with the skeleton first found in Dendrobium. Compounds 2-9, 11, 13, and 19 are reported from Dendrobium for the first time, and compounds 14 and 18 are reported for the first time from D. officinale. Compounds 11 and 14 showed moderate DPPH radical scavenging capacity, and compounds 11-14 demonstrated potent ABTS radical scavenging capacity, possessing antioxidant activity.

Optimization of phosphate oxygen isotope pretreatment measurement method based on phosphate in situ enrichment blanket.

Phosphate oxygen isotope analysis is an effective tool for investigating phosphorus migration and transformation in water bodies. Unfortunately, current pretreatment methods for this technology are significantly limited due to their demanding sample amount requirements, complex operation, and limited scope of application. In order to enhance the efficiency of the pretreatment process, hydrated zirconia was synthesized through liquid-phase precipitation. Zeolite, D001 macroporous resin, activated carbon, and ceramsite were chosen as possible candidate materials for loading purposes. The optimal zirconium loading material was identified through a combination of field enrichment and laboratory elution experiments. The ideal in situ enrichment duration, material dosages, and elution time were ascertained using response surface methodology. The findings showed that D001 resin exhibited superior selective adsorption and elution capacity for phosphate. The response surface optimization yielded the optimal parameters for the in situ phosphate-enrichment blanket: a mass of 13 g for zirconium-loaded D001 resin, an enrichment period of 360 min, and an elution period of 853 min. The attainment of a bright yellow Ag3PO4 solid after purification served as proof of the reliability of the optimization method. The obtained results provide a fundamental basis for the preparation and application of phosphate oxygen isotope analysis in freshwater ecosystem.

Study on Chemical Constituents of the Enriched Extracts of Erigeron multiradiatus and Anti-thrombotic Effect on Zebrafish

Background Erigeron multiradiatus (Lindl.) Benth. has been used for years to treat various diseases in Traditional Tibetan Medicine (TTM). Our previous studies have shown that E. multiradiatus extract has important pharmacological effects, such as anti-myocardial ischemia–reperfusion (I/R) injury, anti-inflammatory, and anti-diabetic effects. To date, there have been no reports on using this extract to treat thrombosis. Objectives The aim of this study was to enrich the active components of E. multiradiatus and to further investigate its anti-thrombotic activity. Materials and Methods We used D101 macroporous adsorption resin to enrich the main active substances of E. multiradiatus and performed quantitative determination using ultra-high-performance liquid chromatography with a photodiode array detector (UHPLC-PDA). Furthermore, we investigated the anti-thrombotic effects of E. multiradiatus using three models: an arachidonic acid-induced platelet aggregation thrombotic zebrafish model, a phenylhydrazine-induced red blood cell (RBC)-injuring thrombotic zebrafish model, and a ponatinib-induced vascular endothelial cell (VEC)-injuring thrombotic zebrafish model. Results The yield of active component enrichment of E. multiradiatus was 4.95% using the D101 resin. We accurately determined eight compounds in the extract. Our results showed that these extracts had significant anti-thrombotic effects in the three different zebrafish models. Conclusion The enriched extract of E. multiradiatus could be used as a natural anti-thrombotic agent. Our results will provide a reference for reasonable clinical applications of E. multiradiatus.

The effect of anthocyanin from Dioscorea alata L. after purification, identification on antioxidant capacity in mice.

Increasing findings devote to searching for natural active compositions as additives to ameliorate health status. Anthocyanin, water-soluble natural pigment, has been concerned due to its favorable antioxidant activity. In this study, we purified anthocyanin from Dioscorea alata L., identified its compounds, and evaluated its antioxidant properties. The results indicated that the purity of anthocyanin increased to 39.59 ± 1.56%, 60.18 ± 1.97%, and 81.08 ± 1.97% after purification via AB-8 macroporous resin, Sep-Pak C18 solid phase, and LH-20 Sephadex stepwise. Ultra-performance liquid chromatography tandem mass spectrometer results indicated that paeoniflorin-3,5-O-dihexoside, petunin-3-O-feruloyl-glucoside-5-O-glucoside, cyanidin-3-O-feruloyl glucoside-5-O-glucoside, cyanidin-3-O-sophoroside, and petunin-3,5-O-dihexoside were the major compounds. The purified anthocyanin exhibited stronger antioxidant activity than the unpurified extract and ascorbic acid, whereas weaker than that of cyanidin-3-O-glucoside in general, which was assessed using DPPH, ABTS, and Fe3+ reducing capacity methods. Moreover, the purified anthocyanin increased GSH-Px, total antioxidant capacity, and superoxide dismutase activities and decreased malondialdehyde concentration on serum in mice after administering lipopolysaccharide for 24 h (p < .05). To summarize, the purified anthocyanin boasts more outstanding antioxidant properties than that of crude extracts. These results provide a reference with source of anthocyanin and it is conducive to use Dioscorea alata L. resources.

The Selective Separation of Carnosic Acid and Rosmarinic Acid by Solid-Phase Extraction and Liquid-Liquid Extraction: A Comparative Study.

Rosmarinus officinalis leaves (ROLs) are widely used in the food and cosmetics industries due to their high antioxidant activity and fascinating flavor properties. Carnosic acid (CA) and rosmarinic acid (RA) are regarded as the characteristic antioxidant components of ROLs, and the selective separation of CA and RA remains a significant challenge. In this work, the feasibility of achieving the selective separation of CA and RA from ROLs by solid-phase extraction (SPE) and liquid-liquid extraction (LLE) was studied and compared. The experiments suggested that SPE with CAD-40 macroporous resin as the adsorbent was a good choice for selectively isolating CA from the extracts of ROLs and could produce raw CA with purity levels as high as 76.5%. The LLE with ethyl acetate (EA) as the extraction solvent was more suitable for extracting RA from the diluted extracts of ROLs and could produce raw RA with a purity level of 56.3%. Compared with the reported column chromatography and LLE techniques, the developed SPE-LLE method not only exhibited higher extraction efficiency for CA and RA, but can also produce CA and RA with higher purity.

Resin structure impacts two-component protein adsorption and separation in anion exchange chromatography

The influence of the resin structure, on the competitive binding and separation of a two-component protein mixture with anion exchange resins is evaluated using conalbumin and green fluorescent protein as a model system. Two macroporous resins, one with large open pores and one with smaller pores, are compared to a resin with grafted polymers. Investigations include measurements of single and two-component isotherms, batch uptake kinetics and two-component column breakthrough. On both macroporous resins, the weaker binding protein, conalbumin, is displaced by the stronger binding green fluorescent protein. For the large pore resin, this results in a pronounced overshoot and efficient separation by frontal chromatography. The polymer-grafted resin exhibits superior capacity and kinetics for one-component adsorption, but is unable to achieve separation due to strongly hindered counter-diffusion. Intermediate separation efficiency is obtained with the smaller pore resin. Confocal laser scanning microscopy provides a mechanistic explanation of the underlying intra-particle diffusional phenomena revealing whether unhindered counter-diffusion of the displaced protein can occur or not. This study demonstrates that the resin's intra-particle structure and its effects on diffusional transport are crucial for an efficient separation process. The novelty of this work lies in its comprehensive nature which includes examples of the three most commonly used resin structures: a small pore agarose matrix, a large-pore polymeric matrix, and a polymer grafted resin. Comparison of the protein adsorption properties of these materials provides valuable clues about advantages and disadvantages of each for anion exchange chromatography applications.

Integrated strategy for the screening of cyclooxygenase-2 inhibitors from triterpenoid saponins in Clematis tangutica.

Screening of novel cyclooxygenase-2 (COX-2) inhibitors from complex natural products is not an easy task. To establish an efficient and feasible strategy for screening COX-2 inhibitors from triterpenoid saponins (TPSs) in Clematis tangutica. Taking TPSs in C.tangutica as example, an optimized macroporous resin (MR) method was established for the enrichment of TPSs. High-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOFMS) was performed to establish the phytochemical profiling of TPSs. Molecular docking was performed to predict the ligand-target interactions and discover the active substances. Chemometric techniques were performed to visualize the structure-effect relationships. High-speed countercurrent chromatography and preparative HPLC were performed to prepare the targets. In vitro activity experiment of COX-2 was performed to verify the virtual screening results. TPSs in C.tangutica were well enriched with the recovery rate of (80.22 ± 2.37)%. Thirty-four kinds of TPSs of oleanane type were deduced by HPLC-QTOFMS. Five TPSs of clematangoside C, clematangoside D, clematangoticoside J, hederoside H1 , and hederasaponin B showed stronger binding abilities with COX-2. The structure with more sugar groups at C-28 may be more conducive to the combination with COX-2. Targets were prepared with purities all above 98%. The IC50 values of target TPSs were 6.03 ± 0.24, 12.44 ± 0.15, 9.36 ± 0.19, 4.78 ± 0.13, and 2.59 ± 0.11 μmol/L, respectively. The integrated strategy using MR, HPLC-QTOFMS, molecular docking, chemometrics, target preparation, and in vitro verification was feasible for rapidly screening COX-2 inhibitors from TPSs in C.tangutica.

Recycling preparative isolation of six bicyclol active metabolites from SD rat urine using macroporous resin, offline 2D LPLC/HPLC, and prep-HPLC combined with pharmacodynamic evaluation of two active metabolites

In our previous study, several bicyclol (BIC) metabolites were found to possess higher solubility, security, and efficacy than the parent drug. However, further research can’t be conducted without monomeric metabolites. In the current study, a highly efficient preparative approach for six BIC-active metabolites from Sprague-Dawley (SD) rat urine was developed. First, 1000 mL of urine was purified and concentrated to 50 mL using microporous resin. Second, middle chromatogram isolated (MCI) GEL®CHP20P adsorbent was used to create a low-pressure liquid chromatography (LPLC) column, which was combined with high-performance liquid chromatography (HPLC) to build an offline 2D system to visualize the separation process. Samples were segmented into 25 tubes and merged into three fractions. Then, recycling preparative HPLC was applied to the monomeric preparation to improve the efficiency. The prepared metabolites possessed high purity (greater than98%), and were verified by nuclear magnetic resonance (NMR). Finally, an isoniazid (INH)-induced liver injury zebrafish model was established to evaluate the efficiency of the BIC, M7, and M8 metabolites. The M7 metabolite exhibited a higher efficiency than BIC in histopathology, gene expression, and aminotransferase levels. Consequently, this study provided a strategy that integrating modern analytical techniques to prepare metabolites for discovering high value candidate compounds from biological metabolism.

Citrus peel polyphenols alleviate intestinal inflammation in mice with dextran sulfate sodium-induced acute colitis

Citrus peel polyphenols have possess the distinct anti-inflammatory activities. However, its underlying mechanism on ulcerative colitis have not been elucidated. The aim of this research was to investigate the anti-inflammatory effect and action mechanisms of citrus peel polyphenols. Total citrus peel polyphenols were concentrated using macroporous resins and separated into water-soluble citrus polyphenols and ester-soluble citrus peel polyphenols. These extracts were then gavaged to acute colitis mice induced by dextran sulfate sodium for 14 days using a dose of 300 mg/kg▪bw. High performance liquid chromatography results showed that the extracts contained flavanones, flavonoids, and phenolic acids. Compared to the dextran sulfate sodium group, total citrus peel polyphenols, water-soluble citrus polyphenols, and ester-soluble citrus peel polyphenols significantly ameliorated the severity of colitis symptoms. Additionally, citrus peel polyphenols reduced the activity of myeloperoxidase, lowered secretion of tumor necrosis factor-α and interleukin-6, and increased interleukin-10. Meanwhile, total citrus peel polyphenols, water-soluble citrus polyphenols, and ester-soluble citrus peel polyphenols effectively blocked the activation of the nuclear factor-kappa B. These results demonstrated that citrus peel polyphenols alleviated ulcerative colitis in mice by damping pro-inflammatory cytokine secretion and suppressing the nuclear factor-kappa B pathway activation.

Preparation, development, and scale-up of standardized pentacyclic triterpenoid-rich extract from Centella asiatica (L.) Urb. and study of its wound healing activity

This pilot-scale study of an innovative green extraction method shows increased biomarker content in plant extracts. Moreover, green extraction methods decrease the effects on the environment and human health and promote industrial growth. This study optimized the process conditions to obtain a pentacyclic triterpenoid-rich extract (PRE) from Centella asiatica (L.) Urb., which contains asiatic acid, madecassic acid, asiaticoside, and madecassoside, and evaluated its biological activities. PRE preparation was scaled up from laboratory to pilot scale. In the pilot scale, a combination of microwave-assisted extraction with an irradiation power of 4 kW and an ultrasonic-assisted extraction at 0.55 kW was used for C. asiatica extraction. The total pentacyclic triterpene content was 106.02 mg/g of crude extract. Then, the C. asiatica extract was fractionated by a macroporous resin (Diaion® HP-20). The PRE preparation method used 50% and 75% EtOH fractions. This PRE produced a high content of pentacyclic triterpenoids at 681.12 mg/g of crude extract. It presented a high anti-inflammatory effect with an IC50 value of 23.88 μg/mL for nitric oxide inhibition and induced wound healing processes (proliferation, migration, and collagen synthesis) in human dermal fibroblast cells. The information gained from this study can advance the industrial extraction of physiologically active substances from various plant sources for use as medicines or components of supplemental food and cosmeceutical products.

Optimizing the Extraction and Enrichment of Luteolin from Patrinia villosa and Its Anti-Pseudorabies Virus Activity.

Luteolin from Patrinia villosa exhibits strong antiviral activity. Here, the conditions for extracting and enriching luteolin from P. villosa were optimized. Response surface methodology was used to determine the optimal extraction parameters in terms of reflux time, solvent ratio, extraction temperature, material-to-liquid ratio, and number of extractions. Thereafter, a macroporous resin method was used to enrich luteolin from P. villosa. Finally, the following optimal extraction and enrichment conditions were established: an extraction time of 43.00 min, a methanol/hydrochloric acid solvent ratio of 13:1, an extraction temperature of 77.60 °C, a material/liquid ratio of 1:22, and a total of two extractions. NKA-9 was determined to be the most appropriate resin for enrichment. The ideal adsorption conditions were as follows: a pH of 5.0, a temperature of 25 °C, an initial luteolin concentration of 19.58 µg/mL, a sample loading volume of 2.9 BV, and a sample loading rate of 2 BV/h. The ideal desorption conditions were as follows: distilled water, 30% ethanol and 80% ethanol elution, and 5 BV at a flow rate of 2 BV/h. After optimization, the enrichment recovery rate was 80.06% and the luteolin content increased 3.8-fold. Additionally, the enriched product exhibited a significant inhibitory effect on PRV (Porcine pseudorabies virus) in vitro and in vivo, providing data for developing and applying luteolin from P. villosa.

Ameliorative effect of scopolamine-induced cognitive dysfunction by Fufangmuniziqi formula: The roles of alkaloids, saponins, and flavonoids

Ethnopharmacological relevanceFufangmuniziqi formula (FFMN), a traditional Uyghur medicine used in China, is derived from an ancient Uyghur medical book and consists of 13 herbs. The herbs of FFMN, such as Peganum harmala L., Glycyrrhiza uralensis Fisch., and Nigella glandulifera, have been demonstrated to have acetylcholinesterase (AChE) inhibitory, anti-neuroinflammatory, or antioxidant effects. Therefore, FFMN may have a good anti-Alzheimer's disease (AD) effect, but its specific action and mechanism need to be further proven. Aim of the studyThis study aims to investigate the anti-AD effects of FFMN and the role played by alkaloids, flavonoids, and saponins in anti-AD. Materials and methodsThe alkaloids, flavonoids, and saponins fractions of FFMN were prepared by macroporous resin chromatography. The absorbed ingredients in the drug-containing serum were identified by UPLC⁃Q⁃TOF⁃MS. An AD mouse model was established by intraperitoneal injection of scopolamine (SCO). The role of different fractions of FFMN in the anti-AD process was examined by Morris water maze (MWM), in-vitro cell, and AChE inhibition assay. ResultsA total of 20 ingredients were identified in the serum samples collected after oral administration of FFMN, and seven compounds were selected as candidate active compounds. MWM experiments showed that different fractions of FFMN could significantly improve SCO-induced learning memory impairment in mice. The alkaloids fraction (ALK) regulated cholinergic function by inhibiting AChE activity, activating choline acetyltransferase activity, and protein expression. Flavonoids and saponins were more potent than the ALK in downregulating pro-inflammatory factors or inflammatory mediators, such as TNF-α, MPO, and nitric oxide. Western blot results further confirmed that flavonoids and saponins attenuated neuroinflammation by inhibiting the phosphorylation of IκB and NF-κB p65. This result was also verified by in-vitro cellular assays. FFMN enhanced antioxidant defense by increasing the activity of superoxide dismutase and reducing the production of MDA. Combined with cellular experiments, flavonoids and saponins were proven more protective against oxidative damage. ConclusionFFMN improved cognitive and memory impairment in the SCO-induced AD mouse model. ALK mainly enhanced the function of the cholinergic system. Flavonoid and saponin fractions mainly attenuated neuroinflammation and oxidative stress by modulating the NF-κB pathway. All these findings strongly suggested that the combination of alkaloid, flavonoid, and saponin fractions derived from FFMN is a promising anti-AD agent that deserves further development.

Isolation, Purification, and Structural Characterization of Polysaccharides from Codonopsis pilosula and Their Anti-Tumor Bioactivity by Immunomodulation.

The activity of polysaccharides is usually related to molecular weight. The molecular weight of polysaccharides is critical to their immunological effect in cancer therapy. Herein, the Codonopsis polysaccharides of different molecular weights were isolated using ultrafiltration membranes of 60- and 100-wDa molecular weight cut-off to determine the relationship between molecular weight and antitumor activities. First, three water-soluble polysaccharides CPPS-I (<60 wDa), CPPS-II (60-100 wDa), and CPPS-III (>100 wDa) from Codonopsis were isolated and purified using a combination of macroporous adsorption resin chromatography and ultrafiltration. Their structural characteristics were determined through chemical derivatization, GPC, HPLC, FT-IR, and NMR techniques. In vitro experiments indicated that all Codonopsis polysaccharides exhibited significant antitumor activities, with the tumor inhibition rate in the following order: CPPS-II > CPPS-I > CPPS-III. The treatment of CPPS-II exhibited the highest inhibition rate at a high concentration among all groups, which was almost as efficient as that of the DOX·HCL (10 μg/mL) group at 125 μg/mL concentration. Notably, CPPS-II demonstrated the ability to enhance NO secretion and the antitumor ability of macrophages relative to the other two groups of polysaccharides. Finally, in vivo experiments revealed that CPPS-II increased the M1/M2 ratio in immune system regulation and that the tumor inhibition effect of CPPS-II + DOX was superior to that of DOX monotherapy, implying that CPPS-II + DOX played a synergistic role in regulating the immune system function and the direct tumor-killing ability of DOX. Therefore, CPPS-II is expected to be applied as an effective cancer treatment or adjuvant therapy.

Extraction methods and compositions of polyphenols in Shanxi aged vinegar

The extraction, purification, qualification, and quantification of polyphenols (PPs) in vinegar are challenging owing to the complex matrix of vinegar and the specific physicochemical and structural properties of PPs. This study aimed to develop a simple, efficient, low-cost method for enriching and purifying vinegar PPs. The enrichment and purification effects of five solid phase extraction (SPE) columns and five macroporous adsorption resins (MARs) for PPs were compared. The results show that SPE columns were more effective in purifying vinegar PPs than MARs. Among them, the Strata-XA column showed a higher recovery (78.469 ± 0.949%), yield (80.808 ± 2.146%), and purity (86.629 ± 0.978%) than other columns. In total, 48 PPs were identified and quantified using SPE and gas chromatography-mass spectrometry from the SPE column extracts; phenolic acids, such as 4-hydroxyphenyllactic acid, vanillic acid, 4-hydroxycinnamic acid, 4-hydroxybenzoic acid, protocatechuic acid, and 3-(4-Hydroxy-3-methoxyphenyl) propionic acid, occupy a major position in SAV. Furthermore, considering the potential applications of PPs, the concentrates were characterized based on their bioactive properties. They exhibited high total PP, flavonoid, and melanoidin contents and excellent anti-glycosylation and antioxidant activities. These results indicate that the established methodology is a high-efficiency, rapid-extraction, and environment-friendly method for separating and purifying PPs, with broad application prospects in the food, chemical, and cosmetic industries.

Enrichment and qualitative analysis of flavonoid glycosides from the aerial parts of Glycyrrhiza uralensis Fisch

Our previous study found that the aerial parts of Chinese liquorice (Glycyrrhiza uralensis Fisch.) had pharmacological effects against chronic non-bacterial prostatitis in rats, however the pharmacologically active compounds remain unclear. Here, a method based on UPLC-Q-Exactive Orbitrap-MS was established to qualitatively analyse the flavonoid glycosides rich fraction extracted from the aerial part of G. uralensis Fisch., after pretreatment with n-butanol and enrichment using AB-8 macroporous resin. Using both positive and negative ion modes, 52 compounds were identified or tentatively characterised by comparison with standards and the literature: 40 flavonoids, 8 organic acids, 2 chromones, 1 coumarin, and 1 phenylethanoid glycoside. This study provides not only an approach to enrich flavonoid glycosides but also a methodology for quickly determining the relevant bioactive components in the aerial parts of G. uralensis Fisch.

Optimization of microwave assisted extraction and biological activities of total flavonoids from grape seed using response surface methodology

Aiming to reduce the environmental contamination and resource wastage generated by discarded grape seed, the management of discarded grape seed should be carried out and their potential bioactivities and pharmacological properties ought to be exploited to the fullest extent possible. A Box–Behnken design of response surface methodology was employed to further optimize microwave-assisted extraction setting for grape seed total flavonoids (GSFE). In addition, the extract (GSFE) of total flavonoids obtained were refined through AB-8 macroporous resin for obtaining more pure flavonoid (GSFP) with determination of their in vitro hypoglycemic activity and antioxidant activity. The results indicated that following optimized extraction conditions were achieved: ethanol concentration 50% liquid-solid ratio 21:1 mL/g, microwave power 540 W, microwave time 126 s. Below mentioned conditions, the extraction yield reached an average experimental value (1.979 mg/g), representing 1.69 times higher than that of water extraction (1.17 mg/g). In contrast to GSFE, the purity of GSFE after purification (GSFP) exhibited a high degree of purity at 13.753 mg/g, representing nearly 2.50 times that of GSFE (3.926 mg/g). GSFP possessed high α-glucosidase and α-amylase inhibitory activities as well as DPPH and ABTS+ free radical scavenging activities with an IC50 value of 0.00641, 0.0257, 0.0669 and 0.0708 mg/mL, inferior to that of GSFE (0.0514, 0.186, 0.129 and 0.223 mg/g). Within present work, the microwave technique for extraction of total flavonoids from grape seeds was investigated with an efficient and cost-effective extraction method. Total flavonoids of grape seeds have certain in vitro blood glucose and antioxidant activity, and the purified total flavonoid biological activity has improved, laying the foundation for further study of its pharmacological activity.

Biosynthesis of diisooctyl 2,5-furandicarboxylate by Candida antarctica lipase B (CALB) immobilized on a macroporous epoxy resin.

Diisooctyl 2,5-furandicarboxylate (DEF), an ester derivative of 2,5-furandicarboxylic acid (FDCA, a bio-based platform chemical), resembles the physical and chemical properties of phthalates. Due to its excellent biodegradability, DEF is considered a safer alternative to the hazardous phthalate plasticizers. Although FDCA esters are currently mainly produced by chemical synthesis, the enzymatic synthesis of DEF is a green, promising alternative. The current study investigated the biosynthesis of DEF by Candida antarctica lipase B (CALB) immobilized on macroporous resins. Out of five macroporous resins (NKA-9, LX-1000EP, LX-1000HA, XAD-7HP, and XAD-8) evaluated, the LX-1000EP epoxy resin was identified as the best carrier for CALB, and the XAD-7HP weakly polar resin was identified as the second best. The optimal immobilization conditions were as follows: CALB (500μL) and LX-1000EP (0.1g) were incubated in phosphate butter (20mM, pH 6.0) for 10h at 35°C. The resulting immobilized CALB (EP-CALB) showed an activity of 639U/g in the hydrolysis of p-nitrophenyl acetate, with an immobilization efficiency of 87.8% and an activity recovery rate of 56.4%. Using 0.02g EP-CALB as the catalyst in 10mL toluene, and the molar ratio of 2,5-dimethyl furanediformate (1mmol/mL) and isooctyl alcohol (4mmol/mL) that was 1:4, a DEF conversion rate of 91.3% was achieved after a 24-h incubation at 50°C. EP-CALB had similar thermal stability and organic solvent tolerance compared to Novozym 435, and both were superior to CALB immobilized on the XAD-7HP resin. EP-CALB also exhibited excellent operational stability, with a conversion rate of 52.6% after 10 repeated uses. EP-CALB could be a promising alternative to Novozym 435 in the biomanufacturing of green and safe plasticizers such as DEF.

Immobilization of Lecitase® Ultra for production of flaxseed oil-based diacylglycerols by glycerolysis and its operational stability

Lecitase® Ultra (LU) was immobilized by adsorption on a macroporous resin and characterized. Meanwhile, the obtained immobilized lipase LXTE-1000-LU was applied in diacylglycerol (DAG) preparation, and the glycerolysis catalytic stability was also investigated. At pH 7.0, resin amount 2.5 g, and adsorption time of 2 h, an immobilization efficiency (IE) of 89.98% could be achieved. The storage stability was promoted after immobilization, in which the lipase activity reached more than 85% after 60 days of storage at 4 °C and 25 °C individually compared to its initial state. Furthermore, 41.31% of flaxseed oil-based DAG was obtained after the glycerolysis reaction using LXTE-1000-LU as the catalyst. Under prolonged reaction, the lipase protein content decreased from 17.91 mg/g to 6.99 mg/g, and significant changes in the secondary confirmation of the protein were also observed, leading to the catalytic activity being reduced.