A good chromatographic separation of the constituents in the ethyl acetate extract of J. procumbens was achieved using a reverse-phase column and a gradient elution with a mixture of water and acetonitrile.
Peptide purification was performed on a Gilson HPLC using a reverse-phase C8 semi-preparative column (ACE, advanced chromatography technologies, USA) with a gradient from 5% to 60% acetonitrile in water (both containing 0.001% v/v trifluoroacetic acid).
HPLC separation was achieved using a reverse-phase XBridge C18 3.5 μm 2.1 × 150 mm column (Waters) and a gradient running from 100% methanol to methanol/isopropanol (1 1 v/v) over 50 min at 0.4 mL/min and 40 °C.
Chromatogrpahic separation by RPLC was achieved using a reversed-phase column and a water/acetonitrile mobile phase, containing 0.05% formic acid and using a gradient system.
It is the reason that Hasse model predicts a reverse gradient for C1 and C10+.
Elution with a reverse gradient is also demonstrated at low salt concentrations for these proteins.
Tissues were then rehydrated through a reverse gradient of ethanol solutions.
Amino-acid analysis was performed using a gradient reverse-phase HPLC system, with precolumn derivatization with o-phtaladehyde (Pierce) and 3-mercaptopropionic acid (Sigma), and fluorescence detection [ 65].
Analytic assessments used a reversed-phase, 4.6(250 mm C18 column (Vydac 218TP54) and a linear 0 to 100% gradient of solvent B applied over 100 min at 1 ml/min.
The method used a reversed-phase C18 column with a mobile phase composed of trifluoroacetic acid solution (0.1%, v/v) and acetonitrile in a gradient mode.
