Because the number of variables in SLN preparation

Because the number of variables in SLN preparation is very high and the development of an optimized method needs many experiments, we useda series of screening tests by varying only one factor at a time to reduce thenumber of independent variables and experiments.

Theresults of the screening tests indicated thatthis system was highly influenced by microemulsion temperature duringthe production. We set the temperature 5 0Cabove melting point of the lipid. A decreasein applying temperature can increase particlesize. On the other hand, the upper temperatures can decompose curcumin readily(31).The effect offour different lipids, glycerylmonosterate, cetyl palmitate, stearic acid and beeswax,was studied on particle size(nm),PDI and stability of SLNs (Table 2).

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Cetylpalmitate was chosen due to the smaller particle size and narrow sizedistribution and most stability of SLNs(32).The effect of probe sonication on size, PDI and %LE of Cur-SLNs wasstudied by varying amplitude and sonication time (Table 3). The best resultswere observed in 30% amplitude at 5 min sonication time. Increased in size andPDI was observed when the amplitude of probe sonication increased to 40%.

It was observed that the application of higher amplitude or sonication time caused the formation of unstable SLNs, increasingparticle size and PDI, which was the result of the destruction of surfactantcoatings because of more kinetic energy. We also investigatedinfluence of homogenization time, mechanical stirring intensity (rpm) and timeon the particle size (nm) and PDI of SLNs and fixed them in 8min, 1000rpm and30 min respectively. Homogenization was performed at 10000 rpm for fivedifferent time intervals 2, 5, 8 , 10 and 15 min.

Dcreased in size and PDI was observed when  homogenization time increased  from 2  to  8  min  while at 10 min  both size and PDI wereincreasing. Homogenization is  one  of the  important  strategies of  applying  kinetic energy  to  achieve lower  size  and PDI.  Applying  high kinetic  energy  for longer  time  may lead  to  instability of  formed  lipidic structures  thereby  resulting into  aggregation  and formation  of  larger particles.

  Size  and PDI  were  higher at  2 and 5  min when  compared  to 8 min due to  insufficient homogenization. Therefore, anoptimum duration of homogenization will result in formation of stable particleswith uniform size distribution (Table 4)