Development and Validation of a Stability-Indicating High-Performance Thin-Layer Chromatographic Method for Determination of Pyridostigmine Bromide in the Presence of Its Alkaline-Induced Degradation Product

Abstract

A validated, sensitive, and highly selective stability-indicating high-performance thin-layer chromatographic (HPTLC) method has been adopted for the quantitative determination of pyridostigmine bromide in the presence of its alkaline-induced degradation product and in pharmaceutical formulations. 3-hydroxy-N-methyl pyridinium bromide (3-OH NMP) is the metabolite, impurity, and alkaline-induced degradation product of pyridostigmine bromide (PB). Pyridostigmine bromide and its alkaline-induced degradation product were separated on silica gel HPTLC F254 plates using methanol–ethyl acetate–triethyl amine–glacial acetic acid (9:1:0.5:0.05 by volume) as the developing system followed by scanning of the separated bands at 270 nm over a concentration range of 2–10 μg band−1 with mean percentage recoveries of 99.84% (SD 1.384). The proposed method was successfully applied to the analysis of pyridostigmine bromide both in bulk powder and in pharmaceutical formulation without interference from other dosage form add...