ABSTRACT
The antioxidant activity and the inhibitory potential of α-amylase of lyophilized hydroethanolic extracts of Conocarpus erectus leaves obtained by ultrasonication were determined. The most potent extract was subjected to ultra-high performance liquid chromatography system equipped with mass spectrometer for metabolite identification. The identified metabolites were docked in α-glucosidase to assess their binding mode. The results revealed that 60% ethanolic extract exhibited highest ferric reducing antioxidant power (4.08 ± 0.187 mg TE/g DE) and α-amylase inhibition (IC50 58.20 ± 1.25 µg/mL. The metabolites like ellagic acid, 3-O-methyl ellagic acid, ferujol, 5, 2 Ì-dihydroxy-6,7,8-trimethyl flavone and kaempferol glucoside were identified in the extract and subjected to molecular docking studies regarding α-amylase inhibition. The comparison of binding affinities revealed 3-O-methyl ellagic acid as most effective inhibitor of α-amylase with binding energy of -14.5911 kcal/mol comparable to that of acarbose (-15.7815 kcal/mol). The secondary metabolites identified in the study may be extended further for functional food development with antidiabetic properties.
Se determinó la actividad antioxidante y el potencial inhibidor de la α-amilasa de extractos hidroetanólicos liofilizados de hojas de Conocarpus erectus obtenidos por ultrasónicación. El extracto más potente se sometió a un sistema de cromatografía líquida de ultra alto rendimiento equipado con un espectrómetro de masas para la identificación de metabolitos. Los metabolitos identificados se acoplaron en α-glucosidasa para evaluar su modo de unión. Los resultados revelaron que el extracto etanólico al 60% exhibió el mayor poder antioxidante reductor férrico (4.08 ± 0.187 mg TE/g DE) e inhibición de la α-amilasa (IC50 58.20 ± 1.25 µg/mL. Los metabolitos como el ácido elágico, 3-O-metil elágico ácido, ferujol, 5, 2 Ì-dihidroxi-6,7,8-trimetil flavona y kaempferol glucósido se identificaron en el extracto y se sometieron a estudios de acoplamiento molecular con respecto a la inhibición de la α-amilasa. La comparación de las afinidades de unión reveló 3-O-metil El ácido elágico como inhibidor más eficaz de la α-amilasa con una energía de unión de -14,5911 kcal/mol comparable a la de la acarbosa (-15,7815 kcal/mol). Los metabolitos secundarios identificados en el estudio pueden ampliarse aún más para el desarrollo funcional de alimentos con propiedades antidiabéticas.
Subject(s)
Plant Extracts/chemistry , alpha-Amylases/antagonists & inhibitors , Myrtales/chemistry , Antioxidants/chemistry , Benzopyrans/analysis , In Vitro Techniques , Plant Extracts/pharmacology , Plant Leaves/chemistry , Molecular Docking Simulation , Antioxidants/pharmacologyABSTRACT
To develop a TLC-densitometric method for the determination of barakol content in Senna siamea leaf and flower extracts, and to compare the barakol content in mature leaves, young leaves and young flowers of the plant which are consumed as a vegetable in curry. The extraction of pure barakol was performed by boiling the fresh young leaves of S. siamea with 0.5% sulfuric acid followed by chloroform extraction. The extract was further purified and recrystallized from absolute ethanol. Authentic sample of barakol was used for the validation of the TLC-densitometric method. Chromatography was performed on a TLC aluminium plate precoated with silica gel 60 F[254] as a stationary phase and chloroform-methanol [85:15 v/v] as a solvent system. Fifteen percent ethanolic extracts of mature leaves, young leaves and flowers of S. siamea were analyzed and compared for barakol content using the validated TLC-densitometric method. Both the validation and analysis of barakol by TLC-densitometry were carried out at the absorbance mode of 366 nm. Barakol was extracted as pure lemon-yellow crystals from young S. siamea leaves with 0.1% yield. Linearity was found over the range of 200-900 ng/spot [r2 = 0.997]. The developed method gave high precision [%RSD < 0.50] and accuracy [average 101.12%]. The limit of detection and limit of quantitation were 8 and 50 ng, respectively. Barakol content in young leaves, mature leaves and young flowers were 1.67, 0.78 and 1.43% dry weight, respectively. R[f] value of the barakol in young leaves, young flowers and authentic sample was the same: 0.45 +/- 0.03. The TLC-densitometric method was simple, precise and convenient; hence it is an effective procedure for the simultaneous determination of barakol in plant extracts
Subject(s)
Plant Leaves/chemistry , Phenalenes/analysis , Flowers/chemistry , Densitometry/methods , Benzopyrans/analysis , Chromatography, Thin Layer/methodsABSTRACT
Senna siamea (Lam.) Irwin and Barneby is a medicinal plant popularly used in Thailand. Young leaves and/or young flowers of this plant have been consumed by Thai people as a Khi Lek curry for a long time. The fresh young leaves and flowers are boiled with water 2-3 times to get rid of the bitterness and the boiled mush is used for curry cooking. Barakol, a major constituent of Senna siamea leaves was analyzed for its content in the fresh young leaves, the boiled leaves and the boiled filtrates by a high-performance thin-layer chromatographic method. Fresh young leaves of S. siamea contained 0.4035% w/w barakol. The amount of barakol in the first and second boiled filtrates were 0.2052 and 0.1079% fresh weight, while the first and second boiled leaves samples were 0.1408 and 0.0414% fresh weight, respectively. The results show the process of preparation of Khi Lek curry by boiling S. siamea young leaves twice with water reduced barakol content up to 90% and the content of barakol in boiled leaves used for curry has much less tendency to cause liver toxicity. This may explain the reason why Thai Khi Lek curry has not caused hepatotoxicity, unlike S. siamea leaves consumed as a powdered capsule.