The objective of this work was to develop and validate a rapid and effective chromatographic
procedure for the quantitative analysis of diphenyl diselenide in poly(lactide) (PLA)
nanoparticles via reversed-phase high performance liquid chromatography (RP-HPLC), using a photodiode
array detector (PDA). Chromatographic runs were performed on a RP C18 column (125 mm, 4.0
mm, 5 µm) with a mixture of methanol and acidified water 0.5% (90:10, v/v) as the mobile phase and
a flow rate of 1.0 mL/min in the isocratic mode. Diphenyl diselenide was detected at a wavelength 240
nm. This method was shown to be specific and linear in the range 10-100 µg/mL (r =0.999). The limits of detection and
quantitation were 0.219 and 0.666 µg/mL, respectively. The method was robust for changes in the mobile phase and flow
rate. The precision (intra- and inter-day) was demonstrated because the maximum relative standard deviation was 3.52%.
The mean recovery for diphenyl diselenide was 99.54% (relative standard deviation = 0.44, n = 9). The method was successfully
used to determine the encapsulation efficiency of diphenyl diselenide in PLA nanoparticles.
Keywords: Diphenyl diselenide, encapsulation efficiency, HPLC, polymeric nanoparticles, validation.
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