LC–MS/MS method of Miltefosine (MFS) was developed for the first time in rat plasma.
The method was validated and stability studies were performed.
Lower limit of quantification of MFS for the LC–MS/MS method was 1 ng/mL.
Absolute bioavailability of MFS was 60.33 ± 2.32% in rats.
MFS has slow absorption, long half-life and high apparent volume of distribution.