Development of Analytical Method to analyze the Encapsulation Efficiency of Brazilian Red Propolis Extract encapsulated in NLC

The aim of this study was to obtain separately mass peak of daidzein (DAI), formononetin (FOR), biochanin (BIO) and analyze them in the Brazilian Red Propolis extract (BRP) through to LC-MS (single quadropole). The mobile phase was a gradient of a mixture of purified water + 0.1 % ammonium acetate:acetonitrile, which started with 80:20 (v/v) until 0:100 (v/v) at 17 min. The flow rate was 0.3 mL/min on column Aquity UPLC BEH C18 (100 x 2.10 mm, 1.7 μm particle size). The injection volume was 10 μL with mass detection of 254, 268, 284 for DAI, FOR, BIO, respectively. Their retention time was 3.31; 4.68; 5.75 minutes. The validation was started to determinate linearity, precision and accuracy, according RDC 166/17. The method was able to identify and separate the three important BRP biomarkers that are essential for determination of encapsulation efficiency (~95%) of this extract encapsulated in nanostructured lipid nanocarrier (NLC).