A fast and simple method for GC-MS quantification of galanthamine in Hippeastrum papilio (Ravena) Van Sheepen and assessment of its alkaloid fractions.

BACKGROUND: Galanthamine, an AChE inhibitor marketed for symptomatic treatment of mild to moderate Alzheimer's disease, is produced by both chemical synthesis and extraction from plant of the Amaryllidoideae subfamily. There are a few validated GC-MS and HPLC-MS methods for its quantification in plant material. All of these methods apply multi-step sample preparation procedures including toxic solvents. The assessment of plant material and development of extraction methodologies from new plant sources (e.g. Hippeastrum papilio) requires validation of quantitative analytical methods considering the species alkaloid pattern.

RESULTS: The extraction of galanthamine from H. papilio raw material was optimized selecting 0.072% of HCl water solution as the most effective extractant. An aliquot (1 µL) of the total extract (1 mL) was directly injected into the GC-MS system avoiding any purification steps with toxic solvents. Selectivity, linearity, sensitivity, precision, accuracy, stability and robustness were determined. The LLOQ and LLOD in SIM mode were found at a concentration of 10 ng/mL and 2 ng/mL, respectively. The method was used for assessment of plant raw material, alkaloid fractions and extraction effectiveness.

SIGNIFICANCE: The method describes the fastest and simplest extraction procedure and direct aqueous injection into the GC-MS for quantification of galanthamine in plant material. As compared to the LC-MS, the use of EIMS detector provides important information and advantage in the assessment of alkaloid fractions from new plant sources of galanthamine.