CHROMATOGRAPHIC ISOLATION AND ENZYME INHIBITION OF LUPININE FROM Anabasis salsa
Abstract
The aerial parts of Anabasis salsa were collected in the Karaganda Region during the flowering stage. To optimize alkaloid recovery, the raw material was alkalized with a 30% KOH solution and extracted via maceration using 90% ethanol. Fractionation and purification were performed on a Spot CPC system (Armen Instrument) using a tailored biphasic solvent system composed of methyl tert-butyl ether (MTBE) : acetonitrile (ACN) : n-butanol : water in a 2:2:1:5 (v/v/v/v) ratio. Qualitative and quantitative profiling of the fractions was conducted via HPLC-ESI-QTOF-MS/MS. The AChE inhibitory activity was evaluated using a fluorometric assay with berberine chloride as a positive control. The optimized extraction workflow with preliminary alkalization resulted in a high lupinine content of 6.35% in the crude extract. Liquid-liquid separation via CPC successfully eliminated irreversible adsorption on solid matrices, reducing sample loss. A total of 28 fractions were collected, and purified lupinine was effectively isolated in fraction 24. In biological assays, the isolated lupinine exhibited strong AChE inhibitory capacity with an inhibition value of 47.90%. Notably, this potential surpassed the performance of the reference standard berberine under identical experimental conditions.
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