Characterization of Butanol fraction of Tragia hispida(THB) by High Performance Liquid Chromatography (HPLC)
HPLC was carried out on 1260 Agilent HPLC System, fitted with an Inert
Sustain C18 (4.6 x 150 mm, 5 µm) column. In these assays, as the highest
sPLA2 inhibitory activity was seen with THB, further characterization by
HPLC was only carried out in this fraction. A solution of THB in
methanol (400 ppm) was filtered first through a PTFE membrane filter
(0.22µm) and then through a C18
cartridge filter (Bond Elut C18), and injected (10 µL) to the HPLC
column. The elution was performed with 1% acetic acid and methanol at a
ratio of 70:30 and the flow rate was maintained at 1mL/min throughout
the procedure. The solvent composition was changed linearly after the 5
minutes to reach a composition of 1% acetic acid: methanol (30:70) at
the end of 25 minutes. The chromatogram was monitored at 254 nm.
Recruitment
of patients with acute dengue infection
Blood samples were collected from 31 adult patients, with a duration of
illness of ≤ 4 days from the National Institute of Infectious Diseases,
Sri Lanka following informed written consent. All of them tested
positive by the NS1 rapid antigen test (SD Diagnostics, South Korea).
They were confirmed to have dengue by qRT-PCR as previously described
and were found to be infected with dengue virus serotype 219. Serum was separated and stored at -200C until further use.