ABSTRACT
The anti-proliferative potential of Passiflora mollissima seeds, an underexplored agri-food waste, was investigated in this work by evaluating the molecular changes induced at transcript and metabolite expression levels on HT-29 human colon cancer cells. For this purpose, a pressurized-liquid extract from P. mollissima seeds obtained under optimized conditions was used for the treatment of HT-29 cells and a multi-omics strategy applied, integrating transcriptomics and metabolomics analysis, along with viability and cell cycle assays to study the molecular mechanisms that explain the anti-proliferative activity of this fruit by-product. After treatment for 48 and 72 h, the viability of HT-29 colon cancer cells was markedly affected, whereas minor effects were observed on normal human colon fibroblast cells. The bioactive extract was shown to arrest HT-29 cells in the S and G2/M phases of the cell cycle, which might be mediated by the inactivation of the FAT10 cancer signalling pathway among other genes identified as altered in the transcriptomic analysis. In addition, cellular redox homeostasis, as well as the polyamines pathway and methionine metabolism were found to be affected as suggested from the metabolomics data. Finally, the Foodomics integration enabled the identification of genes, such as MAD2L1, involved in the polyamine and glutathione metabolism, or the inactivation of the NUPR1 transcription factor, that might be related with the alteration of the intracellular ceramide levels in response to endoplasmic reticulum stress.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Cell Survival/drug effects , Passiflora/chemistry , Plant Extracts/pharmacology , Seeds/chemistry , Antineoplastic Agents, Phytogenic/chemistry , Cell Line , Cell Proliferation/drug effects , Glutathione/metabolism , Humans , Metabolomics , Methionine/metabolism , Plant Extracts/chemistry , Polyamines/metabolismABSTRACT
An integrated analytical methodology based on pressurized-liquid extraction (PLE) in two steps, followed by in vitro assays and liquid chromatography/gas chromatography coupled to high-resolution mass spectrometry (HRMS), was developed and applied for the isolation and characterization of potential bioactive metabolites from Passiflora mollissima seeds. PLE was proposed in two sequential steps: 1) recovery of the lipidic fraction using nonpolar solvents, and 2) recovery of the phenolic fraction from the defatted seeds' residue using polar solvents. Cyclohexane was selected as the most suitable extraction solvent for the seeds defatting process (20 min, 100 °C and 100 bar). PLE optimization by response surface methodology was carried out to obtain phenolics-rich extracts with the highest antioxidant activity. Optimal extraction yield (6.58%), total phenolic content (29.99 mg g-1), total flavonoids content (0.94 mg g-1) and antioxidant activity (6.94 mM trolox g-1 and EC50 of 2.66 µg mL-1) were obtained operating at 150 °C with EtOH (100%) as solvent. Untargeted and semi-targeted MS and MS/MS data-mining strategies were successfully implemented for the rapid and comprehensive profiling of the polar and lipidic PLE fractions analysed by UHPLC and GC, respectively, coupled to quadrupole time-of-flight mass spectrometry (q-TOF-MS/MS). Polyphenols-rich extracts from P. mollisima seeds were characterized for the first time applying this approach, showing a broad variety of flavonoids, genuine flavanols (e.g. (epi)fisetinidol) and abundant proanthocyanidins. This application can be considered a successful demonstration of the great potential of the proposed methodology to effectively obtain and characterize complex natural extracts with potential bioactivity, by making use of powerful integrated identification strategies to facilitate the challenging post-acquisition data processing of huge datasets generated by HRMS analysis.
Subject(s)
Chemistry Techniques, Analytical/methods , Gas Chromatography-Mass Spectrometry , Passiflora/chemistry , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Polyphenols/chemistry , Polyphenols/isolation & purification , Seeds/chemistry , Antioxidants/chemistry , Antioxidants/isolation & purification , Flavonoids/analysis , Liquid-Liquid Extraction , Phenols/analysis , Solvents/analysisABSTRACT
Passiflora mollissima (Kunth) L.H. Bailey is an exotic fruit native to South America, known as taxo in Ecuador. This paper characterizes its flavonoid and carotenoid composition and antioxidant capacity and evaluates the effect of the spray-drying process on its phytochemical composition and antioxidant capacity. A total of 18 flavonoid compounds, nine proanthocyanidins and nine flavan-3-ol monomers, were identified and quantified. Glycosides of (epi)-afzelechin stood out as the most abundant flavonoid. Three carotenoids were identified, with ß-carotene having the highest concentration. The DPPH· and ORAC assay methods indicated a high antioxidant capacity. Furthermore, the bioactive content showed a positive and direct correlation with antioxidant capacity. On the other hand, the spray-drying process produced a stable phytochemical composition and antioxidant activity of taxo. These results demonstrate the potential applicability of microencapsulated taxo as a functional ingredient in the food industry.
Subject(s)
Antioxidants/analysis , Carotenoids/analysis , Flavonoids/analysis , Passiflora/chemistry , Phytochemicals/analysis , Fruit/chemistry , Glycosides/analysis , Phenols/analysis , beta Carotene/analysisABSTRACT
Passiflora mollissima (Kunth) L.H.Bailey "tumbo serrano" es una planta nativa peruana, utilizada en medicina tradicional por sus beneficios en la salud y valor nutricional. Presenta compuestos polifenólicos principalmente flavonoides; responsables de la capacidad antioxidante y mayoría de propiedades farmacológicas. Los objetivos del presente trabajo: determinar la actividad antiinflamatoria del extracto etanólico del fruto de Passiflora mollissima (Kunth) L.H.Bailey "tumbo serrano" utilizando el modelo experimental: edema auricular inducido por TPA y su uso como activo biológico en la industria cosmética a través de pruebas de seguridad in vitro por el método Irritection Assay System (potencial de irritación dérmica) y el método HET CAM (potencial de irritación ocular). El extracto etanólico al 20% de Passiflora mollissima (Kunth) L.H.Bailey "tumbo serrano" presentaron actividad antiinflamatoria en las dosis de 500 y 1000 µg. En las pruebas de seguridad in vitro, se observó ligero potencial irritante. Conclusiones, en las condiciones experimentales se demostró que el extracto etanólico de Passiflora mollissima (Kunth) L.H.Bailey "tumbo serrano" presentó actividad antiinflamatoria y seguridad como activo biológico en la industria cosmética.
Subject(s)
Humans , Passiflora , Cosmetic Industry , Anti-Inflammatory Agents , Peru , PhytochemicalsABSTRACT
The odour-active volatiles of curuba fruit (Passiflora mollissima (Kunth) L. H. Bailey) were isolated by solvent assisted flavour evaporation (SAFE). GC-O and GC-MS analyses identified linalool, hexyl acetate, 1,8-cineole, and butyl acetate as key aroma compounds of this fruit. Other odorants relevant because of their contribution to the overall aroma were: 2-methylpropyl acetate, (Z)-3-hexen-1-ol, and (Z)-3-hexenyl acetate. Sulphur compounds, 3-sulfanylhexyl acetate and methional, were reported here for first time as odour-active volatiles in curuba. By HPLC-ESI-MS analyses of glycosidic mixtures and GC-MS analyses of volatiles released enzymatically with a glucosidase, (Z)-3-hexenyl ß-D-glucopyranoside and linalyl ß-D-glucopyranoside were identified as aroma precursors in P. mollissima fruit. Thermal treatment of the glycosidic mixture at native pH of fruit gave furanoid cis- and trans-linalool oxides, as well as, α-terpineol, compounds that exhibit flowery odour notes. Biogenic relationships among odour-active volatiles and their glycosidic precursors were also proposed.
Subject(s)
Flavoring Agents/analysis , Fruit/chemistry , Passiflora/chemistry , Gas Chromatography-Mass Spectrometry , SmellABSTRACT
Introducción: en Colombia hay un gran número de especies exóticas sin explotación con potencial agroindustrial que constituyen una fuente futura de ingresos. El género Passiflora, el más grande de la familia Passifloraceae comprende cerca de 500 especies, ninguna se caracteriza por poseer un alto contenido de compuestos polifenólicos con propiedades antioxidantes; excepto Passiflora mollissima (Kunth) L.H. Bailey (curuba) que es una fruta con alta capacidad reductora. Objetivos: determinar la capacidad que tienen los extractos acuosos de curuba, proveniente de la región de Antioquia, Colombia, para atrapar especies reactivas de oxígeno. Métodos: los contenidos de fenoles, flavonoides y taninos se determinaron por métodos espectrofotométricos. Los ácidos fenólicos se establecieron por high-pressure liquid chromatography. La capacidad para atrapar radicales libres peroxilos por los métodos fluorescentes ORAC (oxygen radical absorbance capacity) hidrofílico y lipofílico. La capacidad para atrapar las especies reactivas de oxígeno total e hidroxilos, se realizó por métodos fluorescentes y los radicales superóxido por espectrofotometría. Resultados: los extractos acuosos de curuba son ricos en polifenoles, especialmente en taninos, flavonoides y ácidos fenólicos. Además, poseen una alta capacidad para atrapar las diversas especies reactivas de oxígeno, especialmente el radical ROO•, con un valor ORAC igual a 108164,9 mmol de Trolox/100 g de pulpa seca, más alto que la mayoría de las frutas y verduras. Conclusiones: el extracto acuoso de curuba tiene un alto potencial nutracéutico, debido al contenido de polifenoles, que inciden directamente en su capacidad para atrapar radicales especies reactivas de oxígeno.
Introduction: Colombia has a large number of exotic species with unexploited agroindustrial potential as a source of future income. Passiflora is the biggest Passifloraceae family, including about 500 species that do not have high content of polyphenolic compounds with antioxidant properties, except for Passiflora mollissima (Kunth) L.H.Bailey (curuba), a fruit with high antioxidant capacity. Objectives: to determine the ability of aqueous extracts to trap reactive oxygen species in curuba fruits from the region of Antioquia, Colombia. Methods: the contents of phenols, flavonoids and tannins were determined by spectrophotometric methods. The phenolic acids were determined by high-pressure liquid chromatography. The ability to trap free peroxyl radicals was measured by hydrophilic and lipophilic fluorescent ORAC (oxygen radical absorbance capacity) methods. The ability to trap total reactive oxygen species and hydroxyls, and superoxide radicals were ascertained by fluorescent methods and by spectrophotometry respectively. Results: aqueous extracts of passion fruit are polyphenol-rich, particularly tannins, flavonoids and phenolic acids. They also have a high capacity to trap the various reactive oxygen species, especially the radical ROO•, with an ORAC value of 108164.9 mmol Trolox/100 g of dry pulp, which is higher than in most fruits and vegetables. Conclusions: the aqueous extract of passion fruit has high nutraceutical potential due to the polyphenol content that directly affects their ability to trap reactive oxygen species radicals.
