Page 11 - PROPOSAL LATIP Zanamivir_Neat 2

(PBS). The intestinal segment will be isolated and cut open. The muscle layer on

the external surface of the segment will be stripped off and the intestinal sheets
will be then mounted to the pins of the diffusion chamber (Harvard Navicyte,

Warner Instruments, USA). The drug solution will be added to the donor
compartment and the same volume of buffer will be add to the receiver site. 95%

O2 and 5% CO2 gas was aerated in each chamber in order to mix each solution

and will be also to maintain the viability of the membrane. Throughout the
experiment, the temperature will be maintained at 37 C. A volume of 0.1 ml

aliquot will be taken from the receiver site at predetermined time intervals over
120 min. The aliquot will be immediately replaced with an equal volume of buffer

solution and then the drugs will be assayed. The apparent permeability coefficient

will be calculate using the following equation (Eq. (1)):

Papp¼FluxX(1/Area)X(1/60)X(1/C0) (1)

Where Papp is referred as apparent permeability coefficient (cm/s), while the flux,
F, is the slope of linear portion of cumulative transport amount to time at the

steady state (pmol/ml). Area refers to the area of diffusion chamber for transport

(fix value), 1.78 cm2, and C0 is the initial drug concentration (pmol/ml). In this
instance, the best microemulsions and nanoemulsion will be identified from

pseudoternary phase diagram study and characterization study will be evaluated.
In addition, 3.0% w/w of captopril in olive oil, an emulsion formed without the

use of glycerol and sucrose ester surfactant will be used as a control.

3.8 Chromatographic Analysis

3.8.1. Chromatographic Condition
Solutions and mobile phases will be freshly prepared before use. The

analytical column is a C18 column. Elution will be obtained by 98% ultrapure
water and 2% acetonitrile (v/v) with a flow rate of 0.5 ml/ min. All analyses will

be carried out under isothermal conditions at 25◦C with UV detection at 230 nm.
The injection volume will be 10μl.

3.8.2 Standard preparation