Trypanocidal Activity of Purified Precocene I by Reverse-Phase High-Performance Liquid Chromatography from Essential Oil of Ageratum houstonianum aerial Parts

Freshly harvested leaves sample of Ageratum houstonionum were dried under
shade and powdered. Leaf sample of A. houstonionum was extracted by process of
hydrodistillation using a Clevenger-type apparatus for the preparation of essential
oil. Extract from A. houstonianum was prepared by dissolving 5 μL of the essential
oil in 10 mL methanol. All the sample was filtered through a Whatman (Maidstone,
England) stainless steel syringe assembly using a 0.22 μm Durapore (Millipore:
Milford, USA) membrane filter. Purification processes via column chromatography,
thin layer chromatography and preparative thin layer chromatography were done.
Reverse phase HPLC analysis was carried out via a Waters HPLC system consisting
of model 510 and 515 pumps, a Rheodyne injector, a Novapak C18 column (250 ×
4.6 mm i.d.; 4 μm), a model 490 E multi-channel detector and Millennium 2010
sata manager. The mobile phase constituents were filtered using a Durapore
0.22 μm membrane filter. The elution was carried out with a linear gradient of
acetonitrile: water (40:60) to pure acetonitrile in 60 min at a flow rate of 1 mL/
min. detection was at 210, 240, 280 and 320 nm.

Author(s): Shaba P*, Pandey NN, Rao JR, Dey S, Mandal BD, Kurade NP, Singh RK, Bhanuprakash V and Chaudary P

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