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Chemical profiling and biological screening of Thymus lotocephalus extracts obtained by supercritical fluid extraction and hydrodistillation
Authors:Patrícia CostaSandra Gonçalves  Clara GrossoPaula B Andrade  Patrícia ValentãoM Gabriela Bernardo-Gil  Anabela Romano
Institution:a Institute for Biotechnology and Bioengineering, Centre of Genomics and Biotechnology (IBB-CGB), Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, Ed. 8, 8005-139 Faro, Portugal
b REQUIMTE/Laboratório de Farmacognosia, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, R. Aníbal Cunha, 164, 4050-047 Porto, Portugal
c Centre for Biological and Chemical Engineering, IBB; DEQB, IST, Av. Rovisco Pais, 1049-001 Lisboa, Portugal
Abstract:Essential oil and extracts from the aerial parts of Thymus lotocephalus were obtained by hydrodistillation (HD) and supercritical fluid extraction (SFE) in two different collectors, respectively. SFE was conducted at 40 °C and a working pressure of 12 or 18 MPa. The chemical profiles were determined using GC-FID and GC-IT-MS. Oxygen-containing monoterpenes were the primary constituents in the essential oil and SFE extracts collected in the second separator, while the extracts obtained in the first separator were predominantly oxygen-containing sesquiterpenes. A large number of compounds were identified by hydrodistillation and, in contrast, the highest extraction yields were obtained using SFE. Linalool (10.43 ± 1.63%) was the main component in essential oil, whereas camphor (7.91 ± 0.84%) and cis-linalool oxide (7.25 ± 1.45%) were the major compounds in the extracts-2nd separator obtained at pressures of 12 and 18 MPa, respectively. Caryophyllene oxide was the primary constituent identified in the extracts-1st separator (4.34 ± 0.51 and 4.41 ± 1.25% obtained at 12 and 18 MPa, respectively). The antioxidant activity was assessed by ORAC and DPPH assays, and the anti-cholinesterase activity was evaluated in vitro using Ellman's method. The essential oil and SFE extracts (first separator) of T. lotocephalus possessed antioxidant activity and strongly inhibited cholinesterases. We also demonstrated that the acetylcholinesterase and butyrylcholinesterase inhibitory activities of the essential oil could be attributed to 1,8-cineole and caryophyllene oxide, respectively.
Keywords:AAPH  2  2&prime  -azobis(2-amidinopropane) dihydrochloride  ACh  acetylcholine  AChE  acetylcholinesterase  AD  Alzheimer's disease  ATCI  acetylthiocholine iodide  BChE  butyrylcholinesterase  BTCI  butyrylthiocholine chloride  ChE  cholinesterase  CO2  carbon dioxide  DTNB  5  5&prime  -dithiobis[2-nitrobenzoic acid]  DPPH  2  2-diphenyl-1-picrylhydrazyl  BHT  butylated hydroxytoluene  SET  single electron transfer  GC-FID  gas chromatography-flame ionisation detector  GC-IT-MS  gas chromatography-ion trap-mass spectrometry  HAT  hydrogen atom transfer  HD  hydrodistillation  ORAC  oxygen radical absorbance capacity  ROS  reactive oxygen species  SFE  supercritical fluid extraction  TE  trolox equivalents  Trolox  6-hydroxy-2  5  7  8-tetramethylchromane-2-carboxylic acid
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