A reversed-phase high-performance liquid chromatography method with pulsed amperometric detection for the determination of glycosides.

We have developed a reversed-phase high-performance liquid chromatography pulsed amperometric detection (RP-HPLC-PAD) method for the determination of glycosides. It is sensitive, repeatable, and selective without the pretreatment step. Ginsenosides were separated completely in 50 min using an water-acetonitrile gradient as the eluent and detected by PAD under NaOH alkaline conditions. The ginsenoside detection limit (S/N=3) was 0.02-0.07 ng and the quantification limit (S/N=10) was 0.1-0.2 ng. The coefficient of linear regression was 0.9984-0.9998 for concentrations between 1 and 50 microg/mL. The intra- and inter-day precision (RSD) was less than 6.35% in Ginseng Radix and Shy-jiun-tzyy-tang extracts. The average recoveries from Ginseng Radix and Shy-jiun-tzyy-tang extracts were 98.19-105.45% and 96.89-102.22%, respectively.