% EE of ZT-loaded NLCs was indirectly determined after centrifugation through measurement of the amount of un-entrapped ZT in the clear supernatant.22 (link) The prepared NLCs were centrifuged at 11,000 rpm for 90 min. The obtained supernatant was filtered through 0.45 Millipore filter (Bedford, MA, USA) of pore size 0.45 μm (Berlin, Germany) and the absorbance of this clear supernatant was measured spectrophotometrically (Ultraviolet-Visible spectrophotometer (Shimadzu, UV-150-02, Sersakusho, Ltd, Kyoto, Japan)) at the predetermined λmax (282 nm) after suitable dilution with 0.1 N HCl of pH 1.2 similar to the standard curve. Plain NLCs were similarly treated and the obtained supernatant was used as a blank. The % EE was determined using equation 1 :
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$$\% EE\, = \,{{Amount\,of\,entrapped\,drug} \over {{\rm{Total}}\,{\rm{amount}}\,{\rm{of}}\,{\rm{drug}}}}\,x\,100$$
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Amount of entrapped drug = Total drug amount – unentrapped drug amount
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$$\% EE\, = \,{{Amount\,of\,entrapped\,drug} \over {{\rm{Total}}\,{\rm{amount}}\,{\rm{of}}\,{\rm{drug}}}}\,x\,100$$
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Amount of entrapped drug = Total drug amount – unentrapped drug amount
