QUANTITATIVE-DETERMINATION OF CUCURBITACINS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY AND HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY

A simple and precise method using High-Performance liquid [HPLC] and High-Performance Thin-Layer Chromatography [HPTLC] was developed for simultaneous determination of the frequently encountered cucurbitacins B (1), D (2), 3-epi-iso-cucurbitacin D (3), E (4) and I (5) as well as cucurbitacin glycosides E (6) and I (7). A reverse phase column [C-18] was used and elution achieved with acetonitrile-water [2:8, solvent A and 45:55, solvent B] with gradient elution analysis. Reverse-phase HPTLC plates were developed with methanol:water (7:3) and normal-phase plates were developed with toluene-ethylacetate (25:75). Developed plates were scanned with Thin-Layer Scanning Instrumentation. A clean-up procedure was perfected before sample loading to both the HPLC and HPTLC.