Magnetoliposomes were obtained using a modified version of the phase-reverse method followed by extrusion at room temperature into a Liposofast device (Avestin, Inc., Ottawa, Canada) through two polycarbonate membrane filters of 0.2-μm pore size, a minimum of nine times both ways [14].
The method of solid phase reverse siRNA transfection was adapted to the 96-well plate format as described elsewhere (Erfle et al., 2008).
For determination of the concentration of CBHs produced in bioreactor cultivations, a phenyl reversed phase method was developed on HPLC, and the purified TeCBH1 -TrCBM-C and C.l.CBH2b were used for generating a standard curve (Additional file 5).
The liposomes were prepared using reverse phase evaporation method and doxorubicin hydrocholoride, a model drug, was loaded using the ammonium sulphate gradient method.
The present study deals with the development of transferosomal gel containing insulin by reverse phase evaporation method for painless insulin delivery for use in the treatment of insulin dependent diabetes mellitus.
Level of encapsulated DTX was measured using a reverse phase HPLC method.
Modified reverse phase evaporation method was presented by Handa et al., and the main benefit of the method is that the liposomes had high encapsulation efficiency (about 80%[33]].
The concentrations of insulin in the supernatant were determined by reverse phase HPLC method, and total amount of insulin released from the nanoparticles was calculated.
Liposomes made by reverse phase evaporation method can be made from numerous lipid formulations and have aqueous volume-to-lipid ratios that are four times higher than hand-shaken liposomes or multilamellar liposomes[19, 20].
The concentration of insulin was determined by reverse phase HPLC method (Agilent 1200, ZORBAX 300 SB-C18 column 150 mm × 4.6 mm, 5 μm, Agilent Technologies Inc., Santa Clara, CA, USA).[37] The mobile phase consisted of a premixed isocratic mixture of 0.2 M sodium sulfate anhydrous solution adjusted to pH 2.3 with phosphoric acid and acetonitrile (73:27, v/v).
Two uniquely different chromatographic methods were developed and yielded the same result for the radiochemical purity - the reverse phase HPLC method and the isocratic ITLC method.
