The blg/D3 complex at a blg (0.1%, w/v):D3 (110 lM) ratio (v/v)
of 1:2 was formed using a procedure previously described
(Diarrassouba et al., 2013). Lyso concentrated at 0.1% (w/v) was
added to the blg/D3 complex using a blg:D3:Lyso ratio (v:v) of2:4:1. The pH of the mixture was adjusted to 7.5 under continuous
nitrogen flow. The resulting ternary system (blg/D3/Lyso) was incubated
at 4 C in the dark for 24 h. The concentration of D3 in the
supernatant was determined by a specific and validated HPLC
method and the encapsulation efficiency (EE) was calculated using
methods described elsewhere