Rutin Content Research Articles

An HPLC Method to Determine Phenolic Compounds of Plant Extracts: Application to Byrsonima crassifolia and Senna alata Leaves

Pharmacognosy Research,2022,14,4,395-404.DOI:10.5530/pres.14.4.58Published:October 2022Type:Original ArticleAuthors:Marília de A Cavalcante, Janyna dos S Oliveira, Mayra S da S Barreto, Lucas P Pinheiro, Patrick de C Cantuária, Wardsson L Borges, Gabriel A da Silva, and Tiago Marcolino de Souza Author(s) affiliations:Marília de A Cavalcante1, Janyna dos S Oliveira1, Mayra S da S Barreto2, Lucas P Pinheiro2, Patrick de C Cantuária3, Wardsson L Borges4,5, Gabriel A da Silva2, Tiago Marcolino de Souza2 1Federal Institute of Amapá, Macapá, BRAZIL. 2State University of Amapá, Macapá, BRAZIL. 3Instituto de Pesquisas Científicas e Tecnológicas do Estado do Amapá, Macapá, BRAZIL. 4Embrapa Amapá, Macapá, BRAZIL. 5Embrapa Agroindústria Tropical, Fortaleza, BRAZIL. Abstract:Background: The Amazonian Region has a variety of medicinal plants with bioactive compounds, whose characterization could present the potential for sustainable development. Objectives: A method for separating, identifying, and quantifying a mixture of nine phenolic compounds (gallic acid, 3-hydroxybenzoic acid, p-coumaric acid, catechin, myricetin, rutin, quercetin, kaempferol, and cyanidin) was developed, validated, and applied to analyze aqueous and hydroethanolic extracts from Byrsonima crassifolia (L.) Kunth and Senna alata (L.) leaves. Materials and Methods: The separation was carried out by HPLC, using a Shim-pack VP-ODS C18 column (5 μm, 150 x 4.6 mm) at 40°C. Detection was performed at 254 nm and separation occurred in 35 min. Results: The optimized method was validated for each of the nine phenolic compounds. The calibration curve for the phenolic compound standards showed suitable linear fitting and exhibited correlation coefficients greater than 0.990. The LOD and LOQ varied between 6.2807 - 14.8851 μg mL-1 and 6.8002 - 16.0071 μg mL-1, respectively. The method was found to be robust for changes of ±2 ml in mobile phase composition. Byrsonima crassifolia aqueous extracts indicated contents of gallic acid, catechin, rutin, and cyanidin whereas hydroethanolic one did not show the first substance. Senna alata aqueous extract presented only 3-hydroxybenzoic acid and rutin whereas myricetin, cyanidin, quercetin, and kaempferol were also identified in the hydroethanolic one. Conclusion: The HPLC method is efficient, precise, accurate, and sensitive to determining phenolic compounds in plant extracts and it is recommended for efficient assays in routine work. Keywords:Bioactive compounds, Chromatographic method, Plant extractsView:PDF (662.62 KB)

Prevention of polycystic ovary syndrome and postmenopausal osteoporosis by inhibiting apoptosis with Shenling Baizhu powder compound.

Shenling Baizhu powder (SBP) has been shown to reverse the abnormal expression of the aromatic hydrocarbon receptor (AHR) mediated by air pollution. Our study aimed to understand the main ingredient of SBP and investigate its action mechanism in preventing polycystic ovary syndrome (POCS) and postmenopausal osteoporosis (PMO). The active ingredients of SBP with the highest binding affinity to AHR were screened using a Chinese medicine database, and their binding mechanism was simulated using molecular dynamics simulation (MDS). Rutin was utilized to treat ovarian granulosa cell lines and osteoblast cell lines. The cell lines were treated with a gradient of rutin concentration (0.01 mmol/L, 0.05 mmol/L and 0.1 mmol/L) to find the optimal drug dose. PCR was used to detect AHR and apoptosis-related proteins, and WB to detect the expression of AHR, caspase-3 and cleaved-caspase-3. Finally, the CCK-8 cell proliferation assay detected the proliferation of cells. We obtained Rutin through the Chinese medicine database, and dynamics simulation determined its binding sites. Ovarian granulosa cell lines and osteoblast cell lines were treated with Rutin. RT-PCR and western blotting revealed that the expression of apoptosis-associated protein Bcl-2 was elevated, and the expression of AHR, Bax, caspase-3 and PARP were decreased. CCK-8 results showed accelerated proliferation in both cell types. Rutin, the main ingredient of SBP compound, works by binding to AHR, which can improve POCS and PMO by inhibiting cell apoptosis and by promoting cell proliferation.

Effect of co-treatment of microwave and exogenous l-phenylalanine on the enrichment of flavonoids in Tartary buckwheat sprouts.

Tartary buckwheat is rich in flavonoids. The application of physical processing technology and exogenous materials treatment can effectively promote grain germination and the accumulation of bioactive secondary metabolites. The content of four flavonoids, the activities of key enzymes (phenylalanine ammonia-lyase (PAL), chalcone isomerase (CHI), flavonol synthase (FLS)) and the expression of key enzyme genes (FtPAL, FtCHI, FtFLS1, FtFLS2) in Tartary buckwheat sprouts treated with microwave and l-phenylalanine (l-Phe) were investigated, and the relationship between them was analyzed to explore the mechanism of promoting flavonoid accumulation, and to provide a theoretical basis for the development of functional Tartary buckwheat sprout food. Germination can promote the synthesis of flavonoids. The contents of chlorogenic acid and rutin in 7-day sprouts increased by 13 420.63% and 225.12% compared with seeds, respectively. Under the best treatment condition T3 (microwave 250 W, 90 s, 2.9 mmol L-1 L-Phe), the specific activities of PAL, CHI and FLS in 5-day-old sprouts increased by 47.84%, 53.04% and 28.02% compared with control check (CK), respectively; and the expression of FtPAL, FtCHI and FtFlS1 increased by 39.84%, 24.78% and 33.72% compared with CK, respectively. Correlation analysis showed that the content of flavonoids in Tartary buckwheat sprouts was significantly positively correlated with the specific activities of key enzymes (P < 0.01) and dynamically correlated with genes related to the synthesis of three enzymes. It suggested that microwave and l-Phe treatment may promote the synthesis of flavonoids by promoting the expression of key enzymes genes in phenylpropane metabolism and controlling the activity of key enzymes in phenylpropane metabolism. © 2022 Society of Chemical Industry.

A simple, cost-effective colorimetric assay for aluminum ions via complexation with the flavonoid rutin

Aluminum has been linked to deleterious health effects with high concentration, chronic exposure, creating a need for innovative detection techniques. Colorimetric assays are an ideal approach since they are simple, cost-effective, and field adaptable. Yet, commercially available colorimetric assays for aluminum are limited since it forms few colored chelation complexes. Flavonoids, a class of polyphenolic compounds, are one of the few examples that create colored aluminum complexes. Aluminum ions (Al3+) are the main constituent in colorimetric assays for flavonoid detection in food or plant samples. Our assay design was based on colorimetric flavonoid assays, where the assay reported herein was optimized. Specifically, the flavonoid rutin concentration and sample-to-rutin volume ratio (295:5 µL) were optimized to detect Al3+ at low µM concentrations in samples. The assay performed comparably, and in some instances better, than those requiring advanced instrumentation and previously reported colorimetric assays, with a linear range (1–8 µM), sensitivity (7.6 nM), limit of detection (79.8 nM), and limit of quantification (266 nM) for Al3+. The colorimetric assay was accurate (99 ≤ 108 ± 4 ≤ 6% Al3+ recovery), precise (low intra- and inter-assay coefficient of variation (CV) of 3.1 ≤ 5.9% and 4.4%, respectively), and selective for Al3+ ions compared to solutions containing a variety of other mono-, di-, and tri-cations at much higher concentrations (10- to 100-fold higher). Lastly, the colorimetric assay was applicable to complex analysis. It was used to generate a chelation curve depicting the Al3+ chelation capacity of sodium alginate, a biologically derived polymer used as a bioink for 3D bioprinting.

Optimization of micropropagation protocol and elicitation of luteolin and rutin biomolecules using phenylalanine and chitosan in root callus of Rumex hastatus D. Don

Rumex hastatus D. Don, a traditional medicinal plant is known for its antitumorous, antiangiogenic and antibacterial potential. HPLC analysis of different plant parts showed that root had the maximum quantity of luteolin and rutin and thus was selected for the establishment of aseptic cultures. Murashige and Skoog’s medium supplemented with 5 µM TDZ proved best for developing greenish-brown and compact calli in 98% root cultures within 30 days. Upon subculturing such callus pieces differentiated an average of 10.50 ± 1.29 shoots per culture. The excised shoots were transferred on MS + 0.1 µM IBA medium where they developed an average of 12.03 ± 2.49 roots per shoot. These plantlets were successfully acclimatized under greenhouse conditions and their genetic fidelity was evaluated employing SRAP and SCoT markers which revealed their high similarity with mother plant proving their true-to-type nature. Interestingly, the in vitro regenerants showed elevated levels of phenols, flavonoids, luteolin and rutin contents and higher antioxidant activities over the mother plant as revealed by phosphomolybdenum and DPPH assays. For elicitation, 100 mg/L phenylalanine was best for enhancing luteolin content (407.18 ± 10.58 μg/g d.w.) up to 17.42-fold over control (22.10 ± 0.57 μg/g d.w.) whereas, 25 mg/L chitosan proved most effective for accumulation of rutin content (1169.07 ± 46.49 μg/g d.w.) up to 2.74-fold over control (312.41 ± 12.42 μg/g d.w.). To the best of our knowledge, this is the first report on micropropagation and elicitation of luteolin and rutin in R. hastatus through root culture. The present investigation focused on selection of an elite plant part, regeneration through in vitro root callus, genetic fidelity analysis, evaluation of secondary metabolites, antioxidant activity and enhancement of luteolin and rutin using elicitors in Rumex hastatus.

Insight into the self-assembly behavior of α-zein by multi-spectroscopic and molecular simulations: An example of combination with the main component of jujube peel pigments – Rutin

Zein has been widely used as a kind of carrier material for its self-assembly capability, while the mechanism of this process is still elusive. Rutin, one of the flavonoids, has been confirmed as the main ingredient of pigments in jujube peels. In this work, the binding mechanism in the zein/rutin complexes has been systematically studied by using multi-spectroscopic methods and molecular simulations. Results have shown that the encapsulation efficiency of complexes has researched the maximum, 67.30 % ± 1.50 %, when the concentration of rutin is 60 μM. Furthermore, the spherical morphology of complexes has been provided by microstructure characterization. Multi-spectroscopic indicated that a static quenching, alongside strong affinity, occurred in the process of interaction. Hydrophobic interaction has been further proven as the main force in zein/rutin complexes from the results of molecular dynamics simulation. This work is vital to fully utilize zein for the delivery of bio-compounds.

Improving antioxidant activity and modifying Tartary buckwheat bran by steam explosion treatment

Steam explosion (SE) is an efficient technology to disrupt the hard structure of cereal bran and improve its biological activities. In this study, different intensities of SE were applied in Tartary buckwheat bran (TBB) to study 1) the changes of bran structures and 2) the release efficiency of polyphenols. Results illustrated SE diminished the contents of free polyphenols in TBB by 6.24–21.65%, while the bound ones were increased by 15.23–120.81%. The in vitro study demonstrated that the scavenging organic free radical DPPH· activity and oxygen radical absorption capacity (1.6 MPa for 60 s) of total polyphenols were significantly improved. Furthermore, the free quercetin contents were increased by 4–6 times and the free rutin contents were decreased by 60–75%. SE made TBB particle size changed, specific surface area increased, thermal stability enhanced, but polyphenol oxidase (PPO) activity decreased. All the results suggested that SE could enhance the antioxidant activity of TBB polyphenols through changing the constituent proportion, structure, thermal stability, and enzyme activity of bran.

Changes in α-Farnesene and Phenolic Metabolism and the Expression of Associated Genes during the Development of Superficial Scald in Two Distinct Pear Cultivars.

Superficial scald is a postharvest physiological disorder that occurs in pear during and after cold storage. In this study, the superficial scald index; α-farnesene and its oxidation products, conjugated trienols (CTols); phenolic content; and the expression of its related genes were investigated in two different pear cultivars, ‘Wujiuxiang’ (Pyrus communis L.) and ‘Yali’ (Pyrus bretschneideri R.), following 115 days of cold storage at 0 °C followed by 7 days of shelf life at 20 °C. The results indicated that the superficial scald occurred after 115 days of cold storage and became more severe during the shelf life of the ‘Wujiuxiang’ pear, whereas no scald was observed in ‘Yali’. The α-farnesene levels increased rapidly at first and then decreased, while the CTols contents increased significantly in ‘Wujiuxiang’ as compared to ‘Yali’, and the expression levels of the genes involved in α-farnesene and CTols metabolism (HMGR1, HMGR2, GSTU7, GPX5, and GPX6), as well as the phenolic synthesis (PAL1, PAL2, C4H1, 4CL2, C3H, and ANR) of the peel, were significantly up-regulated at the onset of the superficial scald. In addition, the relative conductivity and contents of catechin and epicatechin were higher, and the expression level of the laccase gene (LAC7) significantly increased with the development of superficial scald, while lower contents of chlorogenic acid, arbutin, and isorhamnetin-3-3-glucoside, as well as the lower expression levels of a phenolic-synthesis-related gene (C4H3) and polyphenol oxidase genes (PPO1 and PPO5), were noticed in ‘Wujiuxiang’ as compared to ‘Yali’. The results indicated that the onset and progression of superficial scald were associated with the accumulation of CTols, cell membrane breakdown, and higher catechin, epicatechin, and rutin contents, as well as the expression of associated genes of the peels of pear fruit.

Portable Wireless Intelligent Electrochemical Sensor for the Ultrasensitive Detection of Rutin Using Functionalized Black Phosphorene Nanocomposite.

To build a portable and sensitive method for monitoring the concentration of the flavonoid rutin, a new electrochemical sensing procedure was established. By using nitrogen-doped carbonized polymer dots (N-CPDs) anchoring few-layer black phosphorene (N-CPDs@FLBP) 0D-2D heterostructure and gold nanoparticles (AuNPs) as the modifiers, a carbon ionic liquid electrode and a screen-printed electrode (SPE) were used as the substrate electrodes to construct a conventional electrochemical sensor and a portable wireless intelligent electrochemical sensor, respectively. The electrochemical behavior of rutin on the fabricated electrochemical sensors was explored in detail, with the analytical performances investigated. Due to the electroactive groups of rutin, and the specific π-π stacking and cation–π interaction between the nanocomposite with rutin, the electrochemical responses of rutin were greatly enhanced on the AuNPs/N-CPDs@FLBP-modified electrodes. Under the optimal conditions, ultra-sensitive detection of rutin could be realized on AuNPs/N-CPDs@FLBP/SPE with the detection range of 1.0 nmol L−1 to 220.0 μmol L−1 and the detection limit of 0.33 nmol L−1 (S/N = 3). Finally, two kinds of sensors were applied to test the real samples with satisfactory results.

Chemical Characterization of Selected Algae and Cyanobacteria from Bulgaria as Sources of Compounds with Antioxidant Activity

The current research focused on algae from the waters of the Black Sea—Chaetomorpha linum, Ulva intestinalis, Ericaria crinita, and bioreactors—Chlorella spp. and Arthrospira platensis/cyanobacterium/. Pigment content, total phenolic content, and antioxidant capacity were investigated for their use as pharmaceutical, food, and cosmetic ingredients. E. crinita exhibited the highest antioxidant activity by ORAC and HORAC (463.3 µmol TE/g and 463.3 µmol GAE/g) and the highest total content of polyphenols and rutin of the investigated algae. Lower protein content was found in saltwater algae than in freshwater algae. For the first time, biologically active substances from the Bulgarian A. platensis and Chlorella spp., produced in a bioreactor, have been quantitatively identified. A. platensis contained rutin (141.25 mg/100 g), naringenin (42.17 mg/100 g), quercetin (26.74 mg/100 g), kaempferol, and quercetin-3-β-glycoside. Phenolic acids were isolated: neochlorogenic (172.27 mg/100 g) for lyophilized and 5783 mg/100 g for convection-dried A. platensis. It has been found that the protein content in A. platensis (convection dried) and Chlorella spp. (lyophilized) was nearly 54% higher than that of green algae U. intestinalis from Varna. The lyophilization process reduced the protein content of A. platensis samples by almost 20%. The high protein content of convection-dried A. platensis (43.4%) and lyophilized Chlorella spp. (43.7%) identified them as suitable emulsifiers in colloidal and emulsion systems.

Water Extract of Senecio scandens Buch.-Ham Ameliorates Pruritus by Inhibiting MrgprB2 Receptor.

Senecio scandens Buch.-Ham (S. scandens) belongs to the Compositae family. As a Traditional Chinese medicine, S. scandens has been used in China to treat conjunctivitis, mastitis and vaginitis, it also has the function of antibacterial and relieving itching. Water extract of S. scandens (WSS) was prepared and its quality was controlled by HPLC. The antipruritic effects of WSS were evaluated by itch behavioral experiments. The oxazolone and compound 48/80 were induced to mice scratch behavior, scratch was recorded 30 min after sensitization. The relationship between the antipruritic mechanism and MrgprB2 on mast cell was studied by using mast cell-deficient Kit (W-sh) "Sash" mice and MrgprB2-/- mice. The mast cells were observed by toluidine blue staining. In vitro, the effects of WSS on MrgprB2 were studied by calcium imaging; The whole-cell patch clamp method recorded the MrgprB2 mediate voltage-dependent currents in mast cells. The content of rutin (0.012%) and hyperin (0.014%) in the WSS were determined. WSS could ameliorate the pruritus induced by Oxazolone (inhibition was 41.19%, p = 0.004) and compound 48/80 (inhibition was 50.29%, p = 0.001). Meanwhile, WSS could reduce the number of mast cells in mice skin tissue with allergic contact dermatitis (ACD) (p = 0.002) or compound 48/80 (p = 0.013). In addition, WSS could inhibit the calcium influx (1 mg/mL: p = 0.001, 3 mg/mL: p < 0.0001) and the voltage-dependent currents induced by activation of MrgprB2 on mast cell. WSS also attenuated the calcium influx induced by compound 48/80 in HEK293 cells overexpressing MrgprB2/X2. These results showed that WSS could ameliorate pruritus by inhibiting MrgprB2 receptor on mast cells.

Changes of phenolics contents, antioxidant activities, and enzyme activities in pellicles of Juglans sigillata Dode during fruits development

ABSTRACT Phenolic compounds are important components that affect the taste and health-care function of walnuts. And phenolic compounds in walnut (Juglans sigillata Dode. cv. ‘Qianhe 7’) pellicles at three developmental stages were explored to identify their role. The results showed that the total phenolic content, total flavonoid content, ferric reducing antioxidant power, 2, 2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) and 1, 1-diphenyl-2-picrylhydrazyl free radical scavenging activities in pellicles increased gradually during fruits development, and reached the maximum value at maturation period. Syringic acid, gallic acid, catechin, and rutin contents were highly significantly positively correlated with reducing power or highly significantly negatively correlated with IC50 of free radical scavenging activities. Gallic acid and rutin contents were highly significantly positively correlated with phenylalanine ammonia-lyase (PAL) activity. Total phenolic content, syringic acid, catechins, and rutin contents were highly significantly negatively correlated with cinnamate 4-hydroxylase (C4H) and 4-coumarate CoA ligase (4CL) activities. Ferulic acid content was highly significantly positively correlated with polyphenol oxidase (PPO) activity. Overall, PAL, C4H, 4CL and PPO were key enzymes in the metabolism of polyphenols, and they affected the antioxidant capacity of the pellicles by regulating the content of polyphenol during walnut fruit development.

Effects of bioactive substances isolated from Siberian medicinal plants on the lifespan of Caenorhabditis elegans

Medicinal plants are sources of natural antioxidants. Acting as reducing agents, these substances protect the human body against oxidative stress and slow down the aging process. We aimed to study the effects of bioactive substances isolated from medicinal plants on the lifespan of Caenorhabditis elegans L. used as a model organism.&#x0D; High-performance liquid chromatography was applied to isolate bioactive substances from the extracts of callus, suspension, and root cultures of meadowsweet (Filipendula ulmaria L.), ginkgo (Ginkgo biloba L.), Baikal skullcap (Scutellaria baicalensis L.), red clover (Trifolium pretense L.), alfalfa (Medicágo sativa L.), and thyme (Thymus vulgaris L.). Their effect on the lifespan of C. elegans nematodes was determined by counting live nematodes treated with their concentrations of 10, 50, 100, and 200 µmol/L after 61 days of the experiment. The results were recorded using IR spectrometry.&#x0D; The isolated bioactive substances were at least 95% pure. We found that the studied concentrations of trans-cinnamic acid, baicalin, rutin, ursolic acid, and magniferin did not significantly increase the lifespan of the nematodes. Naringenin increased their lifespan by an average of 27.3% during days 8–26. Chlorogenic acid at a concentration of 100 µmol/L increased the lifespan of C. elegans by 27.7%. Ginkgo-based kaempferol and quercetin, as well as red clover-based biochanin A at the concentrations of 200, 10, and 100 µmol/L, respectively, increased the lifespan of the nematodes by 30.6, 41.9, and 45.2%, respectively.&#x0D; The bioactive substances produced from callus, root, and suspension cultures of the above medicinal plants had a positive effect on the lifespan of C. elegans nematodes. This confirms their geroprotective properties and allows them to be used as anti-aging agents.

Inhibition of α-Glucosidase and Pancreatic Lipase Properties of Mitragyna speciosa (Korth.) Havil. (Kratom) Leaves.

Kratom (Mitragyna speciosa (Korth.) Havil.) has been used to reduce blood sugar and lipid profiles in traditional medicine, and mitragynine is a major constituent in kratom leaves. Previous data on the blood sugar and lipid-altering effects of kratom are limited. In this study, phytochemical analyses of mitragynine, 7-hydroxymitragynine, quercetin, and rutin were performed in kratom extracts. The effects on α-glucosidase and pancreatic lipase activities were investigated in kratom extracts and mitragynine. The LC-MS/MS analysis showed that the mitragynine, quercetin, and rutin contents from kratom extracts were different. The ethanol extract exhibited the highest total phenolic content (TPC), total flavonoid content (TFC), and total alkaloid content (TAC). Additionally, compared to methanol and aqueous extracts, the ethanol extract showed the strongest inhibition activity against α-glucosidase and pancreatic lipase. Compared with the anti-diabetic agent acarbose, mitragynine showed the most potent α-glucosidase inhibition, with less potent activity of pancreatic lipase inhibition. Analysis of α-glucosidase and pancreatic lipase kinetics revealed that mitragynine inhibited noncompetitive and competitive effects, respectively. Combining mitragynine with acarbose resulted in a synergistic interaction with α-glucosidase inhibition. These results have established the potential of mitragynine from kratom as a herbal supplement for the treatment and prevention of diabetes mellitus.

Haskap Berry Leaves (Lonicera caerulea L.)-The Favorable Potential of Medical Use.

The presented research evaluates the medical use potential of Lonicera caerulea leaves, which are waste plants in cultivating berries. The study’s screening activity included the leaves of five varieties of Lonicera caerulea: Atut, Duet, Wojtek, Zojka, and Jugana. The microbiological analysis confirmed the safety of using Lonicera caerulea leaves without significant stabilization. Lonicera caerulea leaves standardization was carried out based on the results of the chromatographic analysis, and it showed differences in the contents of active compounds (loganic, chlorogenic and caffeic acids, and rutin), which are attributed to biological activity. For the Lonicera caerulea leaves varieties tested, the differences in the content of total polyphenol content, chlorophylls, and carotenoids were also confirmed. The screening of biological activity of five Lonicera caerulea leaf varieties was carried out concerning the possibility of inhibiting the activity of α-glucosidase, lipase, and hyaluronidase as well, and the antioxidant potential was determined. The defined profile of the biological activity of Lonicera caerulea leaves makes it possible to indicate this raw material as an essential material supporting the prevention and treatment of type II diabetes. However, this research showed that tested enzymes were strongly inhibited by the variety Jugana. The health-promoting potential of Lonicera caerulea leaves was correlated with the highest chlorogenic acid and rutin content in the variety Jugana.

The Content Variation of Four Active Components in Amygdalus persica L. during Different Harvesting Periods

In this study, a quantitative method for the content determination of rutin, 5-O-coumaroylquinic acid methyl ester, chlorogenic acid butyl ester, and kaempferol in Amygdalus persica L. flowers during different harvest periods was established to investigate its various rules and determine the optimal harvesting period. The determination was performed on the XTERRA MS C18 column with a mobile phase consisting of 0.1% formic acid aqueous solution and acetonitrile (gradient elution) at a flow rate of 1.0 mL/min. In combination with other validation data, including precision, stability, and recovery tests, this method demonstrated good reliability and sensitivity. The results showed that the contents of rutin, 5-O-coumaroylquinic acid methyl ester, chlorogenic acid butyl ester, and kaempferol in A. persica flowers during different harvest periods were quite different, and the content in samples at the early blooming stage was the highest. The method is simple, accurate, and rapid for determining the contents of four active ingredients in A. persica flowers.

Degradation kinetics of rutin encapsulated in oil-in-water emulsions: impact of particle size.

Rutin is a natural bioactive flavonoid that is poor in water solubility and chemical stability. Encapsulation can be used to protect bioactive molecules from chemical or physical decomposition during food processing and storage. Thus, the effect of initial particle size on the ability of oil-in-water emulsions to retain rutin during storage was investigated. Rutin was encapsulated in oil-in-water emulsions with different mean surface-weighted diameters: d3,2 =0.56 μm (small), 0.73 μm (medium), and 2.32 μm (large). As expected, the resistance of the emulsions to coalescence and creaming during storage increased as the particle size decreased due to weakening of the colloidal and gravitational forces acting on the droplets. The concentration of rutin in the emulsions decreased during storage (28 days), which was mainly attributed to photodegradation of the flavonoid. The loss of rutin from the emulsions during storage was fitted using a second-order equation. The rutin degradation rate constant k decreased and the half-life t1/2 increased with decreasing droplet size, which was attributed to the stronger encapsulation and light scattering by smaller oil droplets reducing the amount of light that can penetrate into the emulsions. This study has important implications for the design of more efficacious emulsion-based delivery systems for incorporating health-promoting nutraceuticals into foods. © 2022 Society of Chemical Industry.

Definition of chemical markers for Hancornia speciosa Gomes by chemometric analysis based on the chemical composition of extracts, their vasorelaxant effect and α-glucosidase inhibition

Ethnopharmacological relevanceHancornia speciosa Gomes (Apocynaceae) is a tree found in the Brazilian savannah, traditionally used to treat several diseases, including diabetes and hypertension. The anti-hypertensive activity of H. speciosa leaves (HSL) has been demonstrated in different models and is credited to the vasodilator effect and ACE (angiotensin-converting enzyme) inhibition. The hypoglycemic effect of HSL has been also reported. Aim of the studyTo establish correlations between the biological activities elicited by H. speciosa extracts and the contents of their major compounds, aiming to define chemical markers related to the potential antihypertensive and antidiabetic effects of the species. Additionally, it aimed to isolate and characterize the chemical structure of a marker related to the α-glucosidase inhibitory effect. Materials and methodsExtracts of a single batch of H. speciosa leaves were prepared by extraction with distinct solvents (ethanol/water in different proportions; methanol/ethyl acetate), employing percolation or static maceration as extraction techniques, at different time intervals. The contents of chlorogenic acid, rutin and FlavHS (a tri-O-glycoside of quercetin) were quantified by a developed and validated HPLC-PDA method. Bornesitol was determined by HPLC-PDA after derivatization with tosyl chloride, whereas total flavonoids were measured spectrophotometrically. Identification of other constituents in the extracts was performed by UPLC-DAD-ESI-MS/MS analysis. The vasorelaxant activity was assayed in rat aortic rings precontracted with phenylephrine, and α-glucosidase inhibition was tested in vitro. Principal component analysis (PCA) was employed to evaluate the contribution of each marker to the biological responses. Isolation of compound 1 was carried out by column chromatography and structure characterization was accomplished by NMR and UPLC-DAD-ESI-MS/MS analyses. ResultsThe contents of the chemical markers (mean ± s.d. % w/w) varied significantly among the extracts, including total flavonoids (2.68 ± 0.14 to 5.28 ± 0.29), bornesitol (5.11 ± 0.26 to 7.75 ± 0.78), rutin (1.46 ± 0.06 to 1.97 ± 0.02), FlavHS (0.72 ± 0.05 to 0.94 ± 0.14) and chlorogenic acid (0.67 ± 0.09 to 0.91 ± 0.02). All extracts elicited vasorelaxant effect (pIC50 between 4.97 ± 0.22 to 6.48 ± 0.10) and α-glucosidase inhibition (pIC50 between 3.49 ± 0.21 to 4.03 ± 0.10). PCA disclosed positive correlations between the vasorelaxant effect and the contents of chlorogenic acid, rutin, total flavonoids, and FlavHS, whereas a negative correlation was found with bornesitol concentration. No significant correlation between α-glucosidase inhibition and the contents of the above-mentioned compounds was found. On the other hand, PCA carried out with the areas of the ten major peaks from the chromatograms disclosed positive correlations between a peak ascribed to co-eluted triterpenes and α-glucosidase inhibition. A triterpene was isolated and identified as 3-O-β-(3′-R-hydroxy)-hexadecanoil-lupeol. ConclusionAccording to PCA results, the vasorelaxant activity of H. speciosa extracts is related to flavonoids and chlorogenic acid, whereas the α-glucosidase inhibition is associated with lipophilic compounds, including esters of lupeol like 3-O-β-(3′-R-hydroxy)-hexadecanoil-lupeol, described for the first time for the species. These compounds can be selected as chemical markers for the quality control of H. speciosa plant drug and derived extracts.

Deep eutectic solvent on extraction of flavonoid glycosides from Dendrobium officinale and rapid identification with UPLC-triple-TOF/MS

In this study, ultrasonic-assisted deep eutectic solvent (DES) method was used to extract flavonoids from Dendrobium officinale. The mechanism of DESs extraction differences was investigated by physicochemical characterization and molecular dynamics simulation experiments. Moreover, flavonoids from Dendrobium officinale were rapidly identified and quantitatively analyzed by UPLC-Triple-TOF/MS without removing DES. The results showed choline chloride-lactic acid (Chol-Lac) had the best extraction effect among forty groups of DESs, the extraction yield was up to 35.23 ± 2.12 mg/g. It was speculated that the tight hydrogen bond structure was the key to the better extraction ability of Chol-Lac. Meanwhile, the viscosity was the main physical parameter reflecting the hydrogen bond structure of DES. Fourteen flavonoid glycosides were identified, among which the content of rutin (1.94 ± 0.24 mg/g) was the highest. And the results of cluster analysis showed that different DESs had great differences in the extraction of flavonoid glycosides.

Molecular Shield for Protection of Buckwheat Plants from UV-B Radiation.

Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) and common buckwheat (Fagopyrum esculentum Moench) are adapted to growing in harsh conditions of high altitudes. Ultraviolet radiation at high altitudes strongly impacts plant growth and development. Under the influence of ultraviolet radiation, protecting substances are synthesized in plants. The synthesis of UV-B defense metabolites is genetically conditioned, and their quantity depends on the intensity of the ultraviolet radiation to which the plants and plant parts are exposed. These substances include flavonoids, and especially rutin. Other substances with aromatic rings of six carbon atoms have a similar function, including fagopyrin, the metabolite specific for buckwheat. Defensive substances are formed in the leaves and flowers of common and Tartary buckwheat, up to about the same concentration in both species. In comparison, the concentration of rutin in the grain of Tartary buckwheat is much higher than in common buckwheat. Flavonoids also have other functions in plants so that they can protect them from pests and diseases. After crushing the grains, rutin is exposed to contact with the molecules of rutin-degrading enzymes. In an environment with the necessary humidity, rutin is turned into bitter quercetin under the action of rutin-degrading enzymes. This bitterness has a deterrent effect against pests. Moreover, flavonoids have important functions in human nutrition to prevent several chronic diseases, including obesity, cardiovascular diseases, gallstone formation, and hypertension.