Abstract
Cerebral malaria (CM) is a deadly neural disease caused by Plasmodium falciparum, affecting predominantly the immunosuppressed people. A combination of artemether (AM) and lumefantrine (LMF) is indicated as the first-line therapy in CM. Being a UV-inactive molecule, AM is difficult to analyse in large set of samples using a routine approach. Wide usage and analytical challenges with AM-LMF call for developing a simple, robust and economical method for simultaneous estimation of AM and LMF. The current studies endeavour to develop and validate a derivatization-based HPTLC densitometry methodology, wherein the influence of various derivatization parameters on the sensitivity and assay of both the drug(s) has been studied and optimized too. Effective chromatographic separation of AM and LMF was achieved with mobile phase, consisting of n-hexane: ethyl acetate (8:2 v/v), with good linearity in the concentration range 25–800 ng/spot and 10-800ng/spot for AM and LMF, respectively. High degree of accuracy, precision, robustness, ruggedness, and sensitivity were observed during validation studies. Well-resolved peaks along with high recovery of both the molecules were attained from biological and formulation matrices. The aforesaid method was subsequently employed to evaluate the brain biodistribution of the drugs following intranasal administration of their co-loaded lipid-based nanoformulation in C57BL/6 mice. In a nutshell, the developed method showed immense utility in the simultaneous estimation AM and LMF for analysis of large number of in-process samples, lipid-based nanoformulations and biological matrices.
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