15) to cover the range between from 4.3 to 15, usually with step of 1 unit HLB, see Table.
Blends of surfactants HLB of
Sample Span 80 Tween 80 blend
1 100% 0% 4.3
2 93% 7% 5.0
3 84% 16% 6.0
Then, utilize the blends (Table) to prepare emulsion, 10-20% weight of oil phase, in this case 10-20% of weight of FAME I guess, correspond to weight of blends surfactants (Span80/Tween 80) need to be added. Disperse the blend of emulsifier into the oil phase and then added the aqueous phase, test all blends of emulsifier (table). If all the emulsions seen equally well, with not much noticeable difference, then repeat the test using less emulsifier. On the contrary, if all the emulsions are poor and no show no great difference, repeat the test but use higher emulsifier content.
The mean diameter of droplet of emulsion would be a good measurement to find the HLB of FAME, a dilution 1:100 of emulsion on propylen glycol (Levius and Drommond, 1953) or maybe use a Malver size analyzer. Plot mean diameter of droplets vs HLB, the correct HLB tend to smaller mean droplet.
On this step will find the gross HLB, a second series of experiments with HLB gross + - 1 using 0.5 step or lower.