Development of a Rapid HPLC-UV Method for Simultaneous Quantification of Protodioscin and Rutin in White and Green Asparagus Spears

Asparagus (Asparagus officinalis L.) spears are rich in bioactive compounds such as protodioscin, a saponin, and rutin, a flavonoid. Protodioscin and rutin are routinely quantified separately, and an approach permitting simultaneous measurement would significantly improve speed of analysis. We have optimized an extraction procedure and modified a method of high-performance liquid chromatography by coupling to an ultraviolet detector to simultaneously analyze protodioscin and rutin in asparagus extracts. An acidic ethanol solvent was more efficient than methanol, acetonitrile, or water in coextraction of protodioscin and rutin. Protodioscin and rutin were detected at 210 nm, with retention times of 12.6 min and 7.9 min, respectively. The method was validated by high linear correlations between 3.13 and 1000.0 mu g/mL for protodioscin (r(2) = 0.9999), and between 0.3 and 1087.5 mu g/mL for rutin (r(2) = 0.9997). The limit(s) of detection and quantification for protodioscin were 1.6 mu g/mL and 3.13 mu g/mL, respectively, and for rutin 0.2 mu g/mL and 0.3 mu g/mL, respectively. White asparagus spears and the crown of the plants were revealed to be rich sources of protodioscin and contained 2.59 to 10.4 mg/g dry weight. Green asparagus spears, particularly the upper portion, were rich in rutin and contained between 1.51 and 7.29 mg/g dry weight.