Analytical Method Development and Validation of Trimethylazanium containing Anti-leishmanial Phospholipid Drug Miltefosine by Liquid Chromatography Quadruple Tandem Mass Spectrometry (LC-ESI-MS/MS) API-2000

The phosphocholine and phospholipid that is hexadecyl monoester of phosphocholine (alkyl phospholipid) is a miltefosine drug used to treat all types of leishmaniasis. It is a broad-spectrum antimicrobial, anti-leishmanial, anti-cancer phospholipid drug. It is used for the treatment of breast cancer and cutaneous metastasis. This present study of miltefosine was given to develop. It validated the analytical method by LC-MS/MS (API-2000). It will be applied to the estimation of miltefosine from its solubility sample, force degradation sample, dissolution sample, and impurity analysis. The calibration concentrations of miltefosine were 125, 250, 500, 1000, 2000, and 4000 ng/ml which accuracy was 90.903 – 109.077% and reported stability was 90.71- 97.99%, 92.04 – 98.02%, 90.00 – 95.52%, 89.72 – 98.54% for freeze-thaw, short term, benchtop, autosampler stability respectively. The developed method for determining and quantifying miltefosine in the sample was also validated as per the US-FDA and EMA guidelines. The validation parameters found within the specified regulatory limit, hence acceptable. The present method also has a short run time (6.00 min). The method is simple, specific, highly selective, sensitive, and reproducible.