Diaion HP-20 Research Articles

Phytochemical Screening of Genus Primula Species Growing in Georgia and Study of their Antioxidant and Anti-inflammatory Potentials

Background: In traditional medicine, species of the genus Primula L. are used to treat various health conditions, such as eye disorders, respiratory infections, headaches, epilepsy, insomnia, as expectorants, and to promote wound healing. Objective: The goal of this article was to evaluate the antioxidant and anti-inflammatory activities of three species from genus Primula L. growing in Georgia: Primula macrocalyx, Primula woronowii and Primula saguramica. Methods: Initially, fractions containing both aerial and underground parts were air-dried, ground, and extracted with 80% ethanol. The extract was then concentrated by condensation and further dried through freeze-drying. Subsequently, additional chromatographic separations were carried out on Diaion HP-20 using solvents such as water, methanol (50% and 100%), and 100% ethyl acetate to isolate the desired fractions. To identify flavonoids and triterpene glycosides, the study employed thin-layer chromatography (TLC) alongside preliminary phytochemical tests. The antioxidant activity of these species was estimated in vitro by cell-free systems using ABTS and DPPH assays. The extracts' anti-inflammatory properties were evaluated using an ex-vivo cell system that isolated neutrophils. The study examined the extracts' impact on reactive oxygen species (ROS) production in neutrophils stimulated with PMA, as well as their effect on the catalytic activity of myeloperoxidase (MPO), a marker of inflammation in neutrophils. Results: Flavonoids and triterpene glycosides were primarily identified in the 50% and 100% methanol (MeOH) fractions of Primula species through TLC and preliminary phytochemical tests. For each experiment, gallic acid and quercetin served as standards at a concentration of 1 mg/ml, while the tested samples were prepared at concentrations of 5 mg/ml. Based on the IC50 findings, P. w 3 exhibits the most potent antioxidant and anti-inflammatory properties, as evidenced by the following indicators: ABTS - IC50=8.51 ± 0.18; DPPH - IC50=34.57 ± 0.47; PMN - IC50=0.68 ± 0.04; SIEFED - IC50=1.49 ± 0.7; and classical IC50=1.89 ± 0.01. Conclusion: Among the tested fractions, only the fraction of the 3 species prepared in MeOH (50%) showed the best dose-dependent antioxidant and anti-inflammatory activities, especially P.w 3 which is probably related to the high flavonoid content found in this species.

Evaluación de la composición química del extracto alcohólico de Gynoxys cuicochensis cuatrec: identificación de metabolitos y exploración de sus propiedades farmacológicas

Gynoxys cuicochensis Cuatrec., a member of the Asteraceae family, inhabits the Fierro Urco moor in the province of Loja. Despite not being used in traditional medicine, it possesses significant landscape value due to its striking yellow inflorescence. For investigating its chemical composition and potential pharmacological properties, a phytochemical characterization of the plant’s ethanolic extract was conducted. The extract was obtained through static maceration for three days for the initial filtration, followed by one day for the second and third filtrations. Chlorophyll was se- parated using Diaion HP-20 resin as a solid phase packed in separation funnels, and an ethanol:water liquid phase with a concentration gradient ranging from 6:4 to 9:1 for elution. The dechlorophyllized extract was then freeze- dried and fractionated using gravity column chromatography. The obtained fractions were further purified through microcolumn and preparative chromatography. To elucidate the chemical structure of the molecules, nuclear magnetic resonance spectroscopy and electrospray ionization mass spectrometry were employed. Two metabolites were isolated for this study. The first one is a known glycosylated flavonoid called Nicotiflorin, while the second one is a phenolic derivative named 1,3-di-O-trans-feruloylquinic acid, which lacks a precise pharmacological description. This discovery represents an interesting and unique finding for this species, suggesting its potential medicinal use.

Adsorption of extracellular lipase in a packed-bed reactor: an alternative immobilization approach.

In light of the growing demand for novel biocatalysts and enzyme production methods, this study aimed to evaluate the potential of Aspergillus tubingensis for producing lipase under submerged culture investigating the influence of culture time and inducer treatment. Moreover, this study also investigated conditions for the immobilization of A. tubingensis lipase by physical adsorption on styrene-divinylbenzene beads (Diaion HP-20), for these conditions to be applied to an alternative immobilization system with a packed-bed reactor. Furthermore, A. tubingensis lipase and its immobilized derivative were characterized in terms of their optimal ranges of pH and temperature. A. tubingensis was shown to be a good producer of lipase, obviating the need for inducer addition. The enzyme extract had a hydrolytic activity of 23 U mL-1 and achieved better performance in the pH range of 7.5 to 9.0 and in the temperature range of 20 to 50°C. The proposed immobilization system was effective, yielding an immobilized derivative with enhanced hydrolytic activity (35 U g-1), optimum activity over a broader pH range (5.6 to 8.4), and increased tolerance to high temperatures (40 to 60 ℃). This research represents a first step toward lipase production from A. tubingensis under a submerged culture and the development of an alternative immobilization system with a packed-bed reactor. The proposed system holds promise for saving time and resources in future industrial applications.

Development, characterization and application of 3D printed adsorbents for in situ recovery of taxadiene from microbial cultivations

Microbial cell factories are an attractive alternative to produce high-value natural products using sustainable processes. However, product recovery is one of the main challenges to reduce production cost and make these technologies economically interesting. In this work, new resins were formulated to 3D print hydrophobic adsorbents for the recovery of biologics from microbial cultivations. Benzyl methacrylate (BEMA) and butyl methacrylate (BUMA) were selected as functional monomers suitable for the adsorption of hydrophobic compounds. Pore morphology was tailored through the inclusion of pore forming agents (porogens) in the resin. Different porogens and porogen concentrations were evaluated resulting in materials with different porous networks. Sudan 1 and the anticancer drug paclitaxel were employed as model compounds to test the adsorption performance of hydrophobic and terpene molecules onto the developed 3D printed materials. The material with greatest adsorption capacity was obtained using BEMA monomer with 40 % (v/v) porogen (BEMA40). The performance of BEMA40 to recover taxadiene from small-scale (5 mL) Saccharomyces cerevisiae cultivations was tested and compared with commercial Diaion HP-20 beads. Taxadiene titres on BEMA40 (46 ± 2 mg/L) and Diaion HP-20 (54 ± 4 mg/L) were comparable, with no taxadiene detected in the cells and cell-free media, suggesting near 100 % taxadiene partition on the adsorbents. Compared to commercial beads, 3D printed adsorbents can be customized with adjustments in the resin formulation, are well adaptable to diverse bioreactor types, do not clog sampling ports and columns and are easier to handle during post processing. The results of this work demonstrate the potential of 3D printing to fabricate hydrophobic interaction adsorbent materials and their application in the recovery of biological products.

Adsorption and desorption strategy for recovering β-carotene from crude palm oil: Kinetics, equilibrium and thermodynamics studies

Palm oil uses are widespread in several industry segments. During refining process, minor compounds such as carotenoids, phytosterols, squalene and tocotrienols are lost. The β-carotene is lost due to the bleaching and the high temperature in the deodorization step. The aim of this work is to recover β-carotene before its degradation in the refining process, using adsorption-desorption strategy. Batch adsorption studies were evaluated for five adsorbents: Diaion HP20, Sepabeads SP70, Sepabeads SP700, Diaion HP2MGL, Amberlite XAD7HP. Modeling of kinetics and equilibrium curves were performed for the Sepabeads SP70, which had the best performance in adsorption. The best conditions for adsorption were at the concentration of 30% (w/w) oil in isopropyl alcohol (IPA) micelle at 30 °C for 180 min. Thermodynamics parameters of adsorption revealed a favorable, spontaneous and exothermic process associated with physical bonding. The studies performed on adsorbent reutilization proved that the regeneration is effective and allows the material reuse at least for 5 cycles without changing the adsorption capacity. Additionally, desorption was evaluated in single and multiple steps and the final enrichment factor was 3.40 ± 0.21. This process allows the palm oil utilization for edible purposes after the adsorption step while recovering β-carotene based on its physicochemical characterization.

New Megastigmane and Polyphenolic Components of Henna Leaves and Their Tumor-Specific Cytotoxicity on Human Oral Squamous Carcinoma Cell Lines.

Polyphenols have a variety of phenolic hydroxyl and carbonyl functionalities that enable them to scavenge many oxidants, thereby preserving the human redox balance and preventing a number of oxidative stress-related chronic degenerative diseases. In our ongoing investigation of polyphenol-rich plants in search of novel molecules, we resumed the investigation of Lawsonia inermis L. (Lythraceae) or henna, a popular ancient plant with aesthetic and therapeutic benefits. The leaves' 70% aq acetone extract was fractionated on a Diaion HP-20 column with different ratios of H2O/an organic solvent. Multistep gel chromatographic fractionation and HPLC purification of the Diaion 75% aq MeOH and MeOH fractions led to a new compound (1) along with tannin-related metabolites, benzoic acid (2), benzyl 6'-O-galloyl-β-D-glucopyranoside (3), and ellagic acid (4), which are first isolated from henna. Repeating the procedures on the Diaion 50% aq MeOH eluate led to the first-time isolation of two O-glucosidic ellagitannins, heterophylliin A (5), and gemin D (6), in addition to four known C-glycosidic ellagitannins, lythracin D (7), pedunculagin (8), flosin B (9), and lagerstroemin (10). The compound structures were determined through intensive spectroscopic investigations, including HRESIMS, 1D (1H and 13C) and 2D (1H-1H COSY, HSQC, HMBC, and NOESY) NMR, UV, [α]D, and CD experiments. The new structure of 1 was determined to be a megastigmane glucoside gallate; its biosynthesis from gallic acid and a β-ionone, a degradative product of the common metabolite β-carotin, was highlighted. Cytotoxicity investigations of the abundant ellagitannins revealed that lythracin D2 (7) and pedunculagin (8) are obviously more cytotoxic (tumor specificity = 2.3 and 2.8, respectively) toward oral squamous cell carcinoma cell lines (HSC-2, HSC-4, and Ca9-22) than normal human oral cells (HGF, HPC, and HPLF). In summary, Lawsonia inermis is a rich source of anti-oral cancer ellagitannins. Also, the several discovered polyphenolics highlighted here emphasize the numerous biological benefits of henna and encourage further clinical studies to profit from their antioxidant properties against oxidative stress-related disorders.

Biological production of epicoccamide-aglycone and its cytotoxicity

Epicoccamide (EPC) is an O-d-mannosylated acyltetramic acid of Epicoccum origin and is a bolaamphiphilic fungal polyketide. EPC displays weak toxicity against Staphylococcus aureus and HeLa cell lines. The EPC biosynthetic gene cluster was previously identified in Epicoccum nigrum and knockout of the glycosyltransferase gene (epcB) abolished EPC production. EPC-aglycone was expected in the epcB knockout but was not found. This study demonstrates that extractive culture using the hydrophobic resin Diaion HP-20 resulted in the production of EPC-aglycone, which was isolated using chromatographic separation techniques, and its structural identity was substantiated by chemical analyses. EPC-aglycone displayed strong antibacterial activity against Staphylococcus aureus, with the minimal inhibitory concentration of 1 μg/mL (64 μg/mL for EPC). EPC-aglycone displayed higher levels of growth inhibition against HeLa cell line (the half inhibitory concentration, 19 μM) and WI-38 (15 μM) cell line than EPC (76 μM and 38 μM vs. HeLa and WI-38, respectively). The dose–response curve fit of growth inhibition indicated that EPC-aglycone adopted a shallow curve (low slope factor), which was different from that of EPC, suggesting that their cellular targets are distinct from each other. This study substantiates that the d-mannose attachment is the final step in EPC biosynthesis, showcasing a glycosylation-mediated modulation of the biological activity of simple acyltetramic acid. This study also highlights the usefulness of extractive cultures in mining cryptic microbial natural products.

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.

Chemical composition and cardiotropic activity of Ziziphora clinopodioides subsp. bungeana (Juz.) Rech.f.

Ethnopharmacological relevanceZiziphora clinopodioides subsp. bungeana (Juz.) Rech.f. is a subshrub that is widely distributed in China, Kazakhstan, Kyrgyzstan, Mongolia, Russia, Tajikistan, Turkmenistan, and Uzbekistan. The species is used in traditional medicine for the relief of symptoms connected to cardiovascular diseases like coronary heart disease or hypertension. Aim of the studywas to validate traditional use of Z. clinopodioides subsp. bungeana for the treatment of coronary hearth diseases using in vivo models and to find active compounds responsible for the activity. Materials and methodsMultiple extracts were obtained from the aerial parts of Z. clinopodioides subsp. bungeana using maceration, liquid-liquid extraction, CO2 extraction and ultrasound-assisted extraction. Preliminary screening studies for the evaluation of the efficacy of Z. clinopodioides subsp. bungeana extracts on the model of hemic hypoxia were performed. The most effective samples were selected and included in the main study. Stage 2 of the study evaluated the cardiotropic activity of the selected extracts on a model of chronic heart failure. Preparations were administered to animals intragastrically once a day for 28 days. For the isolation of individual compounds plant material was extracted with 96% ethanol. The obtained crude extract was sequentially extracted with n-hexane and dichloromethane and separated by chromatography on a Diaion HP-20 column. The obtained fractions were further subjected to Sephadex LH-20 column chromatography and eluted isocratically with 96% ethanol (EtOH) to yield subfractions, which were further separated by preparative HPLC to obtain 13 individual compounds. ResultsExtracts obtained from Ziziphora clinopodioides subsp. bungeana (Juz.) Rech.f. herb were subjected to pharmacological screening for the evaluation of their efficacy on hemic hypoxia. Based on the obtained results, out of the sixteen tested extracts two (AR and US 60%) were selected for further evaluation of their cardiotropic activity. Modeling of chronic heart failure was carried out in accordance with the following stages: 1) anesthesia with chloral hydrate at a dose of 450 mg/kg, intraperitoneally, 2) artificial ventilation of the lungs, 3) thoracotomy, 4) modeling of permanent ischemic or ischemic-reperfusion damage. Both extracts effected the indicators of contraction and output, comparable to the reference drug - Monopril. Based on the extraction methods used to obtain RAF and US60 and data from the literature, it can be assumed that they contain compounds with medium polarity, including polyphenols and terpenoids. At the next stage three previously undescribed monoterpenoid derivatives – Ziziphoric acid (1), Ziziphoroside D (2) and 6′-malonylziziphoroside A (3), along with two previously described megastigmane glucosides – blumenol C glucoside (4), blumenol C 9-O-(6′-O-malonyl-beta-D-glucopyranoside (5) and two previously described monoterpenoids 7a-hydroxymintlactone (6), 7-hydroxypiperitone (7) together with six polyphenols – pinocembrine-7-O-rutinoside (8), chrysine-7-O-rutinoside (9), acacetin-7-O-rutinoside (10), luteolin-7-O-rutinoside (11), rutin (12) and rosmarinic acid (13) were isolated from Z. clinopodioides subsp. bungeana extracts. ConclusionOur results support the traditional use of Z. clinopodioides subsp. bungeana for the treatment of coronary diseases. As a result of Z. clinopodioides subsp. bungeana extracts screening in vivo, two extracts were selected as potential cardiotropic agents. Phytochemical analysis of the plant material led to the isolation of five terpenoid derivatives, two megastigmane glycosides, five flavonoids and one cinnamic acid derivative, which could be responsible for the reported biological activity. Future experiments are required to understand the mechanisms of action for the isolated compounds.

Meliasendanins E-J, Nor-neolignan Constituents from Melia toosendan and their Anti-inflammatory Activity

A phytochemical investigation of the fruits extract of Melia toosendan afforded the isolation of two new nor-neolignans, meliasendanins E (1) and F (2), as well as twelve known compounds (3 - 14) using various separation technique such as Diaion HP20, silica, RP-18 gel column chromatography and semi-preparative HPLC. Their chemical structures were elucidated by extensive NMR spectroscopic data including 2D-NMR, and HR-ESI-MS as well as ECD data. Among the twelve known compounds, the absolute structures of 3 - 6 were determined first, and given the trivial names as meliasendanins G-J (3 - 6). Based on the evaluation of anti-inflammatory activity, compounds 7 - 8 exhibited inhibitory effects on LPS-induced nitric oxide production in RAW 264.7 macrophages with IC50 values of 34.6 and 39.5 μM, respectively.

Comparison of Four Different Polymeric Adsorbents as Clean-up Agents for Pesticide Analysis in Tea with LC MS/MS

Previously published studies indicate that tea (Camellia sinensis) contains essential oils, flavonoids, phenolics, lipids, natural pigments, sugars, and oligosaccharides. These essential biomolecules in Camellia sinensis cause serious problems in the determination of pesticide residues. High amount of co extractives can exhibit chromatographic characteristics similar to some pesticides, which may cause serious matrix effects and significant interference in mass spectrometry analysis. In this study, we compared four polymeric resins as clean-up sorbents for the determination of 216 pesticide residues in tea by a high performance liquid chromatography triple quadrupole mass spectrometry. styrene-divynilbenzene (Diaion HP20), Polyamide 6, Polyvinylpolypyrrolidone (PPVP), and Amberlite XAD 7 were used as clean-up sorbents instead of PSA. However,to evaluate the effectiveness of the adsorbents, recovery studies were carried out using these adsorbents. In addition, GC-MS analysis was also performed to see the ability of these adsorbents to remove co-extracts. According to the recovery results, when the acceptable limit was accepted as 60-140%, it was determined that the resin that gave the best results with 170 pesticides was polyamide. Diaion was the second resin with 166 pesticides, while XAD, PSA which was used as control group and PPVP were determined as the third, fourth, and fifth resins with very close values of 159, 155, and 152 pesticides, respectively. According to GC-MS total ion analysis, it was observed that PSA gave the chromatogram with the least co- extract, while it was observed that PSA was the worst adsorbent in removing caffeine, one of the main bioactive compounds in tea. XAD-7 and Polyamide were found to be the best adsorbents in removing caffeine.

Bio-Fabrication of ZnONPs from Alkalescent Nucleoside Antibiotic to Control Rice Blast: Impact on Pathogen (Magnaporthe grisea) and Host (Rice).

In the traditional method of the bio-fabrication of zinc oxide nanoparticles (ZnONPs), bacterial strains face metal toxicity and antimicrobial action. In the current study, an alkalescent nucleoside antibiotic was mixed with zinc hexanitrate to fabricate the ZnONPs. An integrated approach of DIAION HP-20 macroporous resin and sephadex LH-20 column chromatography was adopted to separate and purify alkalescent nucleoside AN03 from Streptomyces koyanogensis. Alkalescent nucleoside was confirmed by the Doskochilova solvent system. The bio-fabricated ZnONPs were characterized by using Fourier transform infrared (FTIR), X-ray diffraction (XRD), and transmission electron microscopy (TEM) analyses. The XRD spectrum and the TEM images confirmed the crystallinity and the spherical shape of the ZnONPs with an average size of 22 nm. FTIR analysis showed the presence of functional groups, which confirmed the bio-fabrication of ZnONPs from alkalescent nucleoside ANO3. In-vitro studies showed that 75 μg/mL of ZnONPs had a strong inhibitory zone (28.39 mm) against the Magnaporthe grisea and significantly suppressed the spore germination. SEM and TEM observations respectively revealed that ZnONPs caused breakage in hyphae and could damage the cells of M. grisea. Greenhouse experiments revealed that the foliar spray of ZnONPs could control the rice blast disease by 98%. Results also revealed that ZnONPs had positive effects on the growth of the rice plant. The present study suggested that ZnONPs could be fabricated from microbe-derived nucleoside antibiotics without facing the problems of metal toxicity and antimicrobial action, thus overcoming the problem of pathogen resistance. This could be a potent biocontrol agent in rice blast disease management.

Production and bioprocessing of camptothecin from Aspergillus terreus, an endophyte of Cestrum parqui, restoring their biosynthetic potency by Citrus limonum peel extracts

The metabolic potency of fungi as camptothecin producer elevates their prospective use as an industrial platform for commercial production, however, the loss of camptothecin productivity by fungi with the storage and subculturing are the major obstacle. Thus, screening for endophytic fungal isolates inhabiting ethnopharmacological plants with an obvious metabolic stability and sustainability for camptothecin biosynthesis could be one of the most feasible paradigms. Aspergillus terreus ON908494.1, an endophyte of Cestrum parqui was morphologically and molecularly verified, displaying the most potent camptothecin biosynthetic potency. The chemical identity of A. terreus camptothecin was confirmed from the HPLC, FTIR and LC–MS/MS analyses, gave the same molecular structure and mass fragmentation patterns of authentic one. The purified putative camptothecin displayed a strong anticancer activity towards HepG-2 and MCF-7 with IC50 values 0.96 and 1.4 µM, respectively, with no toxicity to OEC normal cells. As well as, the purified camptothecin displayed a significant antifungal activity towards fungal human pathogen Candida albicans, Aspergillus flavus, and A. parasiticus, ensuring the unique structural activity relationships of A. terreus camptothecin, as a powerful dually active anticancer and antimicrobial agent. The camptothecin productivity of A. terreus was maximized by bioprocessing with Plackett–Burman design, with an overall 1.5 folds increment (170.5 µg/L), comparing to control culture. So, the optimal medium components for maximum yield of camptothecin by A. terreus was acid why (2.0 mL/L), Diaion HP20 (2.0 g/L), Amberlite XAD (2.0 g/L), dextrin (5.0 g/L), glucose (10.0 g/L), salicylic acid (2.0 g/L), serine (4.0 g/L), cysteine (4.0 g/L) and glutamate (10.0 g/L), at pH 6 for 15 days incubation. By the 5th generation of A. terreus, the camptothecin yield was reduced by 60%, comparing to zero culture. Interestingly, the productivity of camptothecin by A. terreus has been completely restored and over increased (210 µg/L), comparing to the 3rd generation A. terreus (90 µg/L) upon addition of methanolic extracts of Citrus limonum peels, revealing the presence of some chemical signals that triggers the camptothecin biosynthetic machinery. The feasibility of complete restoring of camptothecin biosynthetic-machinery of A. terreus for stable and sustainable production of camptothecin, pave the way for using this fungal isolate as new platform for scaling-up the camptothecin production.

Synthesis of hexyl butyrate (apple and citrus aroma) by Candida rugosa lipase immobilized on Diaion HP-20 using the Box-Behnken design.

The online version contains supplementary material available at 10.1007/s10068-022-01200-1.

Antiglycation Effects of Adlay Seed and Its Active Polyphenol Compounds: An In Vitro Study.

This study aimed to evaluate the antiglycation effects of adlay on protein glycation using in vitro glycation assays. Adlay seed was divided into the following four parts: the hull (AH), testa (AT), bran (AB), and polished adlay (PA). A solvent extraction technique and column chromatography were utilized to investigate the active fractions and components of adlay. Based on a BSA-glucose assay, the ethanolic extracts of AT (ATE) and AB (ABE) revealed a greater capacity to inhibit protein glycation. ATE was further consecutively partitioned into four solvent fractions with n-hexane, ethyl acetate (ATE-Ea), 1-butanol (ATE-BuOH), and water. ATE-BuOH and -Ea show marked inhibition of glucose-mediated glycation. Medium–high polarity subfractions eluted from ATE-BuOH below 50% methanol with Diaion HP-20, ATE-BuOH-c to -f, exhibited superior antiglycation activity, with a maximum inhibitory percentage of 88%. Two phenolic compounds, chlorogenic acid and ferulic acid, identified in ATE-BuOH with HPLC, exhibited potent inhibition of the individual stage of protein glycation and its subsequent crosslinking, as evaluated by the BSA-glucose assay, BS-methylglyoxal (MGO) assay, and G.K. peptide-ribose assay. In conclusion, this study demonstrated the antiglycation properties of ATE in vitro that suggest a beneficial effect in targeting hyperglycemia-mediated protein modification.

Phytochemical characterization and anti-inflammatory activity of a water extract of Gentiana purpurea roots

Ethnopharmacological relevanceGentiana purpurea was one of the most important medicinal plants in Norway during the 18th and 19th centuries, and the roots were used against different types of gastrointestinal and airway diseases. Aim of the studyTo explore the content of bioactive compounds in a water extract from the roots, a preparation commonly used in traditional medicine in Norway, to assess the anti-inflammatory potential, and furthermore to quantify the major bitter compounds in both roots and leaves. Materials and methodsG. purpurea roots were boiled in water, the water extract applied on a Diaion HP20 column and further fractionated with Sephadex LH20, reverse phase C18 and normal phase silica gel to obtain the low molecular compounds. 1D NMR, 2D NMR, and ESI-MS were used for structure elucidation. HPLC-DAD analysis was used for quantification. The inhibition of TNF-α secretion in ConA stimulated peripheral blood mononuclear cells (PBMCs) was investigated. ResultsEleven compounds were isolated and identified from the hot water extract of G. purpurea roots. Gentiopicrin, amarogentin, erythrocentaurin and gentiogenal showed dose-dependent inhibition of TNF-α secretion. Gentiopicrin is the major secondary metabolite in the roots, while sweroside dominates in the leaves. ConclusionsThe present work gives a comprehensive overview of the major low-molecular weight compounds in the water extracts of G. purpurea, including metabolites produced during the decoction process, and show new anti-inflammatory activities for the native bitter compounds as well as the metabolites produced during preparation of the crude drug.

Use of Diaion HP20 Resin to Achieve High Rutin Containing Moringa oleifera Extract: Its Solubility and Anti-Bacterial Properties and Possible Applications in Nebulizer Formulation for COVID Patients

Rutin with several pharmacological properties has been reported to be an effective inhibitor for SARS-COV-2 viral protease. Due to lack of specific drugs available for treatment of covid infection, hunt is on for possible herbal supplements that will impact multiplication of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and also provide immunity to people against such infections. We report a simple and cost-effective method for extraction of rutin from Moringa oleifera, a plant with high nutritive value due to its rich contents in minerals, vitamins and other essential phytochemicals. The solubility of rutin in water was achieved using rutin-arginine mixture (identified as a good water-soluble version of rutin) and tested for its antibacterial activity against Klebsiella pneumoniae and E. coli by MIC studies and compared with rutin solubilized in methanol. Our results demonstrate easy scale of the rutin extraction process and such a process could be applicable to extraction of rutin from other medicinal plants as well. We demonstrate that rutin purified through column chromatography has 20% higher solubility in water and maintains its anti-bacterial properties against Klebsiella pneumoniae and E. coli. Our findings reveal new possibilities of using resin column chromatography for concentrating rutin from plant extracts. Also, potential use of water soluble rutin is envisaged for development of nebulizers for treatment of asthma, hyperglycemia, and pneumonia, seen in COVID-19 affected patients.

Chemical and biological assessment of high value pharmaceutical raw material from Egyptian orange peel

In Egypt, orange juice industries generate huge amount of waste peel that could be a source of raw materials with high value and economic return. Here, we explored a better model for waste management of citrus processing waste in Egypt by developing an ecofriendly method for preparation of value added materials. A high grade pectin (HGP) was obtained from the crude acidic water extract of the peel after passing through Diaion HP20 column. The HGP showed potent antidiabetic activity at lower dose than those reported in literature. It possessed significant effect on blood glucose (BG) level, as well as parameters relevant to liver and kidney functions in streptozotocin (STZ) induced diabetic rats. On the other hand, the essential oil (EO) prepared by cold press showed the highest yield (0.72% w/w) and it is the most applicable method of isolate orange oil on pilot scale. EO showed significant antimicrobial activity against the tested food borne pathogens. In conclusion, high value materials — HGP and EO were prepared on pilot scale from the waste orange peel. While the HGP can be included in food supplement for diabetic patients, EO can be used as a natural food preservative.

Lactic acid-based deep eutectic solvent: An efficient green media for the selective extraction of steroidal saponins from Trillium govanianum

Natural deep eutectic solvents have a high solvation and food-grade characteristics, which make them a preffered choice for extracting bioactives to develop nutraceutical and cosmetic formulations. This research aimed to investigate the systematic effects of several NADESs on the ultrasonic-assisted extraction of steroidal saponins from the rhizomes of Trillium govanianum Wall. ex D.Don, which is utilized in traditional Ayurvedic medicine. After screening of fifteen different NADESs, choline chloride: lactic acid (1:1) (NADES 1) was found most effective eutectic solvent for the extraction of steroidal saponins performed by the three-round micro-extraction (1 mL). Compared with hydro-ethanolic solvent-based extraction, lactic acid-derived NADESs showed higher extractability of steroidal saponins. The extraction efficiency of bioactive steroidal saponins e.g. borassoside E and protodioscin was also enhanced with NADES 1. The effects of extraction variables such as S:L ratio, working duration, and temperature on the extraction of steroidal saponins by NADES 1 was also studied and optimized.Furthermore, optimized parameters were validated by performing 100 mL NADES 1-based extraction to recover phytomolecules and NADES 1 by using Diaion HP20 resin column chromatography. As a proof of concept, this study suggested that lactic acid-derived NADESs are effective media for the extraction of steroidal saponins from biomass. The cytocompatibility assay of NADESs with NRK-52E and IEC-6 cells revealed the safe nature of NADESs to normal animal cells. The steroidal saponins-enriched extract obtained with NADES 1 has shown promising antagonistic effects on acetylcholinesterase activity.

Effective Utilization of Vaccinium virgatum Aiton Stems as Functional Materials: Major Constituent Analysis and Bioactivity Evaluation.

We previously reported that rabbit-eye blueberry (Vaccinium virgatum Aiton) leaves exhibit multiple functions. In this study, we evaluated whether V. virgatum stems can also be used as functional materials similar to leaves and clarified the major constituents and their biological activity (antioxidant activity and anti–adult T cell leukemia (ATL) activity). Water extracts of V. virgatum stems were separated into 19 fractions using a Diaion HP-20 open column. Sugars and organic acids were detected in the highly water-soluble fractions. Polyphenols and proanthocyanidin were detected in the hydrous methanol-soluble fractions. In biological activity evaluations, a difference in antioxidant activity was observed in the water-containing methanol-eluted fractions, and fractions exhibiting anti-ATL activity differed depending on cell type. These results suggest that blueberry stems, like leaves, are rich in polyphenols and exhibit antioxidant activity and inhibit ATL cell growth. In the future, aerial parts of blueberries, including stems and leaves, could be used as functional materials and/or medicinal resources.