The effect of concentration on the DST of DPPC in PBS with Ca2+ was also studied. At 1000 ppm concentration, DPPC dispersions produced low tension minima (γmin), lower than 10 mN/m, after 10 minutes pulsation, which is the fastest compared with other concentrations. The γmin also reached to the value, lower than 10 mN/m, after 20 minutes pulsation at pulsating rate of 20 rpm right after generating bubble.
The effect on the ability of DPPC to lower the DSTs in water, PBS, PBS with Ca2+, and HEPES with Ca2+ was also studied with another method of preparing the DPPC dispersions, devised by Bangham and others. The DPPC dispersions prepared with the Bangham method can produce low tension minima (γmin), lower than 10 mN/m, in all conditions except with PBS solutions. The time it takes to reach DSTs below the 10 mN/m limit is longer than with the new method by 10 minutes. The effect on the DPPC vesicle size was also examined. The diameter of the DPPC vesicles is larger than with the new method, and especially in HEPES with Ca2+, the diameter is 980 nm whereas the one prepared with new method is 70 nm. In addition, while the new method's DPPC vesicles remained about the same for one month, the other method's vesicles increased day by day. The results show the dispersions for the new method are much more stable. These results have implications for designing efficient protocols of lipid dispersion preparation and for effective administration of lung surfactant replacement formulations for treating alveolar respiratory diseases.