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An ultra-high-performance liquid chromatographic (UHPLC) separation was developed for six kava pyrones (methysticin, dihydromethysticin (DHM), kavain, dihydrokavain (DHK), desmethoxyyangonin (DMY), and yangonin), two unidentified components, and three Flavokavains (Flavokavain A, B, and C) in Piper methysticum (kava). The six major kavalactones and three flavokavains are completely separated (Rs > 1.5) within 15 min using a HSS T3 column and a mobile phase at 60 °C. All the peaks in the LC chromatogram of kava extract or standard solutions were structurally confirmed by LC-UV-MS/MS. The degradations of yangonin and flavokavains were observed among the method development. The degradation products were identified as cis-isomerization by MS/MS spectra. The isomerization was prevented or limited by sample preparation in a non-alcoholic solvent or with no water. The method uses the six kava pyrones and three flavokavains as external standards. The quantitative calibration curves are linear, covering a range of 0.5â»75 µg/mL for the six kava pyrones and 0.05â»7.5 µg/mL for the three flavokavains. The quantitation limits for methysticin, DHM, kavain, DHK, DMY, and yangonin are approximately 0.454, 0.480, 0.277, 0.686, 0.189, and 0.422 µg/mL. The limit of quantification (LOQs) of the three flavokavains are about 0.270, 0.062, and 0.303 µg/mL for flavokavain C (FKC), flavokavain A (FKA), and flavokavain B (FKB). The average recoveries at three different levels are 99.0â»102.3% for kavalactones (KLs) and 98.1â»102.9% for flavokavains (FKs). This study demonstrates that the method of analysis offers convenience and adequate sensitivity for determining methysticin, DHM, kavain, DHK, yangonin, DMY, FKA, FKB, and FKC in kava raw materials (root and CO2 extract) and finished products (dry-filled capsule and tablet).
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Flavonoides/química , Kava/química , Lactonas/química , Extractos Vegetales/química , Dióxido de Carbono/química , Cromatografía Líquida de Alta Presión/métodos , Isomerismo , Límite de Detección , Estructura Molecular , Raíces de Plantas/química , Piranos/química , Pironas/química , Relación Estructura-Actividad , Espectrometría de Masas en Tándem/métodosRESUMEN
Guayusa tea is derived from the leaves of the Ilex guayusa Loes. plant, which is native to the Amazon rainforest. Beyond its pleasant sensory properties, Guayusa tea is rich in antioxidants, phenolics, and minerals. In this study, the effects of infusion time, temperature, and solvent conditions on the color, antioxidant capacity, total phenolic content, phenolic profile, and antimicrobial activity of Guayusa (Ilex guayusa Loes.) tea were investigated. Guayusa tea samples were prepared using two different solvents, ethanol and water, with 4, 6, and 8-h infusions at 60 and 70 °C. Liquid chromatography, diode array detection, and electrospray ionization tandem mass spectrometry (LC-DAD-ESI-MS/MS) were used to determine a comprehensive profile of phenolic compounds and to detect differences due to infusion conditions. Moreover, after the Guayusa tea infusion with the highest bioactive properties was determined, the effects of in vitro gastrointestinal digestion on the total phenolic content, antioxidant capacity, and phenolic compounds of the Guayusa tea infusion were measured. Phenolic profile analysis identified 29 compounds, among which chlorogenic acid and its derivatives were predominant. The increase in infusion time was correlated with an elevation in total phenolic content. Significant differences were observed between water and ethanol infusions of Guayusa in terms of phenolics and antioxidants. The total amount of phenolic compounds in the samples prepared with both solvents was found to increase after oral intake, depending on the digestion stage; meanwhile, the amounts of flavonoid compounds and di-O-caffeoylquinic acid derivatives decreased during digestion.
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In the original publication [...].
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Liver and muscle health are intimately connected. Nutritional strategies that support liver detoxification are beneficial to muscle recovery. Computational-in silico-molecular systems' biology analysis of supplementation of calcium and potassium glucarate salts and their metabolite D-glucaric acid (GA) reveals their positive effect on mitigation of liver detoxification via four specific molecular pathways: (1) ROS production, (2) deconjugation, (3) apoptosis of hepatocytes, and (4) ß-glucuronidase synthesis. GA improves liver detoxification by downregulating hepatocyte apoptosis, reducing glucuronide deconjugates levels, reducing ROS production, and inhibiting ß-Glucuronidase enzyme that reduces re-absorption of toxins in hepatocytes. Results from this in silico study provide an integrative molecular mechanistic systems explanation for the mitigation of liver toxicity by GA.
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Ácido Glucárico , Biología de Sistemas , Especies Reactivas de Oxígeno , Músculos/metabolismo , Glucuronidasa/metabolismo , Hígado/metabolismoRESUMEN
According to the Food and Agriculture Organization (FAO), protein demand is expected to increase globally by around 40% by 2030 as a response to the world's population growth. Due to their clean label, vegan or vegetarian based applications, nutritional value, and cost-efficient properties, plant-based proteins have been widely studied. However, most of the alternatives currently found in the market have some challenges because of their poor solubility, emulsifying, gelling, and foaming attributes. Hemp seed protein has gained increasing attention due to its unique amino acids and fatty acids profiles. In this study, commercial HPC mixtures were adjusted to pH 2, 4, 6, 8, 10, and 12 followed by ultrasonication (US) for 5 min (5 s on: 5 s off) and incubated for an hour before neutralizing to pH 7. Following the treatments, the samples were analyzed for their hydrodynamic diameter, conductivity, zeta potential, polydispersity index, surface hydrophobicity, solubility, electrophoresis (SDS-PAGE), free sulfhydryl group, and optical characteristics. The samples treated with ultrasound at pH 8 and 10 significantly (p < 0.05) enhanced the solubility of the hemp seed protein by 12.12% and 19.05%, respectively. Similarly, the samples treated with ultrasonication and pH shifting at pH 6, 8, and 10 also significantly increased the amount of free sulfhydryl content (p < 0.05) to 41.6, 58.72, and 46.54 mmol/g from 32.8 mmol/g, respectively. This study shows that the application of ultrasonication and pH shifting is a promising alternative method to modify the functional properties of HPC and widen their applications in the food, cosmetics, and pharmaceutical industries.
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ETHNOPHARMACOLOGICAL RELEVANCE: Kava and kava extracts have shown great potential as a way to minimize anxiety-associated symptoms and to help alleviate pain. Hepatoxicity has been associated with the consumption of kava products. The chemical compounds, kavalactones (KL) and flavokavains (FK) have been implicated in kava's psychotropic and possible hepatotoxic properties. AIM OF THE STUDY: To investigate the kavalactone and flavokavain content and in vitro toxicity of KAVOA™, a supercritical carbon dioxide extraction (SFE) of kava. MATERIALS AND METHODS: Kavalactone and flavokavain content of SFE kava and noble kava root were determined following extraction in acetone, cell culture media, and water using ultra high-performance liquid chromatography (UHPLC). Using water extractions of the kava products, the cell viability and toxicity on the human hepatocellular carcinoma cell line (HepG2) were determined using luminescent and fluorescent assays, respectively. The half maximal inhibitory concentration (IC50) of the SFE kava and noble kava root, extracted in cell culture media, were determined utilizing a luminescent cell viability assay. RESULTS: Quantification of the KAVOA™, a SFE extract of kava and kava root showed similar profiles of kavalactone and flavokavain content. Water extracted SFE and root kava did not show a negative impact on cell viability and toxicity when compared to the vehicle control treated cells. IC50 values were determined for the SFE kava and kava root extracted in cell culture media in respect to cell viability, 78.63 and 47.65⯵g/mL, respectively. CONCLUSIONS: KAVOA™, a supercritical carbon dioxide extract of kava displays a similar kavalactone profile to a noble variety of kava. In relation to total kavalactone content, KAVOA™ also has a lower content of the cytotoxic compound FKB. Aqueous extractions of KAVOA™ and noble kava root had no significant negative impact on cell viability and toxicity on HepG2 cells when compared to vehicle controlled treated cells. Results indicate KAVOA™ demonstrates a similar in vitro safety profile to that of noble kava root when experiments are normalized to kavalactone content.
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Flavonoides/aislamiento & purificación , Kava/química , Lactonas/aislamiento & purificación , Extractos Vegetales/toxicidad , Carcinoma Hepatocelular/metabolismo , Supervivencia Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión/métodos , Cromatografía con Fluido Supercrítico , Flavonoides/química , Células Hep G2 , Humanos , Concentración 50 Inhibidora , Lactonas/química , Neoplasias Hepáticas/metabolismo , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Pruebas de Toxicidad/métodosRESUMEN
The aim of this study was to test the performance of a HPLC method, designated for rapid quantification of chlorogenic acids (CGA) in green coffee extract (GCE). The precision statistics associated with the method were assessed using three independent laboratories with five samples analyzed in triplicate. Seven main CGA isomers (3-CQA, 5-CQA, 4-CQA, 5-FQA, 3,4-diCQA, 3,5-diCQA and 4,5-diCQA) were quantified. The concentration of total CGA in the samples varied from 32.24% to 52.65% w/w. The repeatability and reproducibility standard deviations for the determination of individual isomers varied, respectively, from 0.01 to 0.28 and 0.05-1.59. The repeatability and reproducibility standard deviations of the calculated total CGA, corresponding to the sum of the seven main CGA isomers, varied respectively, from 0.17 to 0.58 and 0.55-2.01. The fast HPLC method evaluated in this study was considered precise and appropriate for the determination of CGA in GCE.
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Café , Ácido Clorogénico , Cromatografía Líquida de Alta Presión , Extractos Vegetales , Reproducibilidad de los ResultadosRESUMEN
A gas chromatography-mass spectrometry (GC-MS) method using the standard addition methodology was developed for the determination of glucuronolactone (GL) and glucuronic acid (DGuA) in four beverages categorized as detoxification, recovery, or energy drinks. The method features a precolumn derivatization step with a combination of BSTFA (N,O-bis(trimethylsilyl)trifluoroacetamide) and TMCS (trimethylchlorosilane) to silylate the analytes. The sample pretreatment required no extraction, filtration, or reduction step prior to the injection. The quantification of the analytes was performed using a five-point standard addition protocol. The proposed method presented excellent intraday precision (%RSD < 10) and linearity for GL calibration curves (correlation coefficients > 0.995) and acceptable linearity for DGuA calibration curves (correlation coefficients > 0.97). The estimated limits of detection (LOD) and quantification (LOQ) for GL ranged from 0.006 ppm to 0.14 ppm, and 0.02 ppm to 0.47 ppm, respectively. The estimated LOD and LOQ for DGuA determination ranged, respectively, from 0.06 ppm to 1.1 ppm and 0.2 ppm to 3.8 ppm. The results demonstrated that the method should be regarded as a reliable alternative to the simultaneous determination of GL and DGuA.