Abstract: HPLC method was applied for analysis of main constituents of 10 lots different growing period samples. The HPLC column, mobile phase elution mode (isocratic or gradient) and gradient program were optimized in order to obtain high quality HPLC profile. The HPLC system consisted of Waters 600 pump and a 996 photodiode-array detector (DAD). HPLC analysis was performed on a Planetsil C18 column (5 μm, 200 mm×4. 6 mm) and Phenomenex HPLC Guard Cartridge System ODS-C18(4×3. 0 mm ID) with the mixture of methanol (A) and H2O (acidified to pH 2. 0 with phosphoric acid) (B) as mobile phase in gradient mode. Concentrations of A 30%, 100%during 0, 50min, column temperature 40℃, detection wavelength 254 nm. The HPLC chromatographic fingerprinting of main constituents showing 27 common peaks was established from 10 lots of samples. These peaks were divided into 3 groups: from 0 to 17 min, 14 peaks (including peak 1~14);from 17 to 35min, 10 peaks (including peak 15~24);from 35 to 50 min, 3 peaks (including peak 25~27). This method with high specificity and good reproducibility can provide scientific basis for determining the best collecting time, quality evaluation and identification of Rumex gmelini.

Key words: Rumex gmelini, HPLC, fingerprinting