The amount of encapsulated CLO per mg of dry LS was determined by disgregating 50 mg of LS in 5 mL of ethanol under stirring (300 r.p.m.) at 60 °C for 2 h.
The samples were filtered (nylon membrane filters, 0.2 μm pore size, Merck Millipore, Milan, Italy) and analyzed by high-performance liquid chromatography (HPLC) for CLO content, as previously reported [19 (link)]. HPLC determinations were performed using a two-plungers alternative pump (Jasco, Tokyo, Japan), an UV-detector operating at 210 nm, and a 7125 Rheodyne injection valve with a 50 μL loop. The samples were loaded on a stainless steel C-18 reverse-phase column (15 × 0.46 cm) packed with 5 μm particles (Hypersil BDS, Alltech, Fresno, CA, USA).
The elution was performed with a mobile phase containing methanol/water 80:20 v/v at a flow rate of 0.8 mL/min. The retention time of CLO was 6.8 min.
CLO encapsulation efficiency (EE) was calculated as follows [11 (link)]:
All data were the mean of four determinations on different batches of the same type of LS.
The samples were filtered (nylon membrane filters, 0.2 μm pore size, Merck Millipore, Milan, Italy) and analyzed by high-performance liquid chromatography (HPLC) for CLO content, as previously reported [19 (link)]. HPLC determinations were performed using a two-plungers alternative pump (Jasco, Tokyo, Japan), an UV-detector operating at 210 nm, and a 7125 Rheodyne injection valve with a 50 μL loop. The samples were loaded on a stainless steel C-18 reverse-phase column (15 × 0.46 cm) packed with 5 μm particles (Hypersil BDS, Alltech, Fresno, CA, USA).
The elution was performed with a mobile phase containing methanol/water 80:20 v/v at a flow rate of 0.8 mL/min. The retention time of CLO was 6.8 min.
CLO encapsulation efficiency (EE) was calculated as follows [11 (link)]:
All data were the mean of four determinations on different batches of the same type of LS.