Development and validation of a high-performance thin-layer chromatography densitometric method for the simultaneous determination of novel 1-acridinyl-1,2,3-triazole derivatives

Abstract

High-performance thin-layer chromatographic (HPTLC) method provides a simple, sensitive, and accurate analytical method for the simultaneous determination of certain synthesized 1-acridinyl-1,2,3-triazole derivatives without interference from starting materials and intermediates. Separation was carried out on Merck HPTLC silica gel 60F254 plates, using chloroform‒methanol (9:1, V/V) and hexane‒ethyl acetate (3:2, V/V) as mobile phases. Validation of the method was performed based on the basis of the International Council for Harmonisation (ICH) guidelines in terms of linearity, sensitivity, limit of detection, limit of quantification, precision, selectivity, and specificity. Least-square equations were calculated for the studied compounds in the ranges of 25–500 and 10–500 ng/spot for ultraviolet (UV) and fluorescence measurements, respectively. Correlation coefficients (r) values were found ranging from 0.9913 to 0.9992 for analytes. The method provides selectivity and specificity which ensure that synthesized compounds are in the pure form without the interference of starting materials and intermediates. The detection limits for the studied compounds ranged from 11.02 to 51.09 ng/spot and 3.84 to 31.95 ng/spot and quantification limits were 33.39–154.82 ng/spot and 11.63–73.67 ng/spot for both spectrophotometric and spectrofluorimetric methods, respectively, indicating applicability for good qualitative and quantitative determination of members of this series at the nanogram concentration levels in biological fluids.