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IJPER · Lens · Bioassay-Guided Isolation and Structure Elucidation of Bioactive Phytoconstituents with Inhibitory Activity against Carbohydrate-Hydrolyzing Enzymes from the Aerial Parts of Premna odorata Blanco

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Article · Volume 55, Issue 3 · pp. 846–856

Bioassay-Guided Isolation and Structure Elucidation of Bioactive Phytoconstituents with Inhibitory Activity against Carbohydrate-Hydrolyzing Enzymes from the Aerial Parts of Premna odorata Blanco

Ronel Abad Mendoza1, Chien-Chang Shen2, Po-Wei Tsai3, Kathlia Aguila De Castro-Cruz1

  1. 1School of Chemical, Biological and Materials Engineering and Sciences, Mapúa University, Metro Manila, PHILIPPINES.
  2. 2National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, TAIWAN.
  3. 3Department of Medical Sciences Industry, College of Health Sciences, Chang Jung Christian University, Tainan, TAIWAN.

Abstract

Background: This study aimed to isolate the bioactive components from the aerial parts of Premna odorata Blanco and to evaluate the hypoglycemic potentials of the crude extracts, sub-fractions and final isolate.Materials and Methods: The plant material underwent a series of enzyme assay-guided isolation and purification having the fractions assayed for inhibitory activity against α-amylase using 3,5-dinitrosalicylic acid (DNS) colorimetric method for each stage. H.1 isolate was tested for α-amylase and α-glucosidase inhibitory activity and glucose uptake by yeast cells. Phytochemical characterization and 1H and 13C NMR spectral analysis were done for the structure elucidation of H.1.Results: In vitro amylase studies revealed that at 100 μg/mL concentrations, hexane crude extract, fraction F (20%/80% and 10%/90% n-hexane/dichloromethane fractions) and F.3 (10:1 v/v petroleum ether/ethyl acetate sub-fraction) exhibited 34.38±0.116%, 71.86± 4.909% and 42.16±1.257% inhibition against α-amylase (at 1000 μg/mL concentration), respectively. H.1 isolate exerted significant inhibition (p < 0.05) of 55.99± 2.202% and 72.43±3.988% against α-amylase and α-glucosidase enzyme (both at 1000 μg/ mL concentration), respectively and significant glucose uptake of 13.85± 0.368%. The purified isolate was spectroscopically confirmed as a 5:1 mixture of β- sitosterol (H.1a) and stigmasterol (H.1b).Conclusion: The compounds have significant inhibitory activity against carbohydrate-hydrolyzing enzymes and may be potentially developed as adjuvant pharmacotherapy for type 2 diabetes.

Keywords: Premna odorata; α-amylase; α-glucosidase; Bioassay; Hyperglycemia; Inhibition

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