Ultrasensitive TLC determination of montlukast and loratadine mixture in human plasma utilizing fluorescence detection at dual pH values: Toward attaining separate maximum fluorescence intensity

Abstract

A novel selective and highly sensitive TLC densitometric method with reflectance/fluorescence detection was developed to separate and quantify montlukast (MONT) and loratadine (LOR). Separation of the studied drugs was performed on precoated silica gel TLC plates using chloroform–ethyl acetate (8 :2 v/v) as a mobile phase. MONT quantification was carried out by measuring emission using 400 nm optical filter after excitation at 340 nm. Enhancement of the week LOR fluorescence was performed through adequate spraying the chromatograms with 0.2 M perchloric acid leading to enhancing sensitivity by 17 folds compared to the reported HPTLC methods with absorbance detection. The scanner was set at 275 nm excitation wavelength and 400 nm optical filter. Detection of both drugs on the same plate separately at different pH conditions was utilized for the first time. Maximum fluorescence was achieved for each of them and this enhances detection sensitivity for both drugs. The linear regression analysis data of the studied drugs produced a good linear relationship with correlation coefficients of 0.996 for MONT and 0.998 for LOR over the concentration range of 6 – 150 ng/band for MONT and 15 – 120 ng/band for LOR. Limit of detection values were 1.7 and 4.5 ng /band for MONT and LOR respectively. The developed method enabled the detection of MONT and LOR in human plasma samples within linear concentrations ranged from 7 to 140 ng/ band and 16 to 50 ng/ band with detection limits of 1.9 and 4.8 ng/band for MONT and LOR respectively.The analytical performance of the proposed method was evaluated according to the International Council for Harmonization (ICH). The method was successfully applied for the analysis of the studied drugs in spiked human plasma and good recovery percentages were obtained indicating that there is no interference from plasma constituents. Therefore the method can be applied for in vivo analysis and pharmacokinetic study.