Rapid TLC and HPLC Quantification of Cucurbitacin C in Cucumber Cotyledons

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Abstract
Two rapid methods for estimating cucurbitacin C (Cuc C) in cucumber cotyledons are described. Both methods require a single cotyledon extracted with methanol. The filtered extract is injected into a C18 HPLC column with methanol-water (55:45) solvent, and quantitative estimates of Cuc C are obtained by UV detection at 254 nm. In the 2nd method, the extract is concentrated for spotting on silica gel TLC plates, the TLC plates are developed with ethylacetate : benzene (75:25), and visualized with vanillin-orthophosphate to give a semiquantitative estimate of Cuc C. The 2 methods are quick and suited for screening seedlings for cucurbitacin levels in segregating populations to select for cucumber beetle resistance and for nonbitter fruit.

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