Page 8 - PROPOSAL LATIP Zanamivir_Neat 2

3.3.1 Particle size analysis

The z-average particle size and particle size distribution of nanoparticles
will be analyze using a Nano C analyzer equipped with a dual 30 mW laser (with

658 nm). Dynamic light scattering (DLS) will be employed to measure the
intensity of the light scattered by the nanoparticle in the suspension at a scattering

angle of 165º. The polydispersity index (PDI) was obtained to represent the size

distribution of the nanoparticle. The physical stability of the nanoparticle will be
monitored by the size change as a function of time.

3.3.2 Polydispersity index (PDI)
Size distribution and polydispersity index of the sample will be measure

by Dynamic Light Scattering (DLS) analysis instrument (Patel et al., 2019).

3.3.3 Morphology study (SEM and TEM)
The morphological characterization study of the sample will be conducted out

using Scanning electron microscopy (SEM) and Transmission electron
microscopy (TEM).

3.3.4 FTIR
The nanoparticles sample will be mixed with potassium bromide (KBr)

crystals at 1:10 ratio and ground using pestle and mortar before being compressed

with hydraulic compressor until it become translucent pallets. The infrared
wavelength of 400 to 4000 nm will be use in the analysis using an Agilent Variant

640 Fourier Transform Infrared spectroscopy.
3.3.5 XRD

The amorphous and crystallization state of the polymers and drug will be

analyze using the Rigaku X-ray Diffraction system (Japan). The instrument
operation will be at 40 kV, 20 mA and the sample will be analyzed at 2º to 60º

angles with angular speed of 5º/min. The scintillation counter slit will be set at
2mm and receiving slit will be opened at 2mm prior to the analysis. The sample

will be place onto 1mm glass slit and distributed evenly prior to analysis.

3.4 Caco-2 cell permeation study

The method for Caco-2 cell permeation study will be based on the method by
(Kamiya et al., 2020) with slight alteration. The Caco-2 cells will be cultured in DMEM

supplemented with nonessential amino acids, penicillin–streptomycin–amphotericin B,