Determination of encapsulation efficiency (EE%) and loading efficiency(LE%) of the nanoparticles

The procedure was followed with suitable changes from (Zhang et al. 2018; Wei et al. 2017) with changes to suit our experiments. CD-1, CD-2 and CD-3 were centrifuged at 10,000 rpm (High Speed Centrifuge, TGL-16B, Shanghai, China) for 20 min leading to the separation of docetaxel and doxorubicin from the NPs. With a 0.45-μm filter, the supernatant was collected.

An HPLC system (Agilent, US) having the following prerequisites was used for the investigation of free drugs in CD-1, CD-2 and CD-3: a TCC 300 C18 column (250 mm × 4.6 mm, 5 μm), a identifying wavelength of 425 nm and a mobile phase of 7.5:7.5:85, v/v of methanol/acetonitrile/0.4% aqueous phosphoric acid and the flow rate of 1 mL/min. After CD-1, CD-2 and CD-3 were dissolved in ethanol; the amount of the total drug loaded was calculated. The multiple drug encapsulation efficiency (EE) along with loading efficiency (LE) was the result of the following formulae:

The equations were employed to decide the EE and LE% of CD-1, CD-2 and CD-3.