Abstract:

Objective To establish a high performance liquid chromatography（HPLC） for the determination of 6 main components in Huang-Lian-Jie-Du decoction. Methods The chromatography was performed on a Zorbax Eclipse Plus C18 column（250 mm×4.6 mm，5 μm） with 0.03 mg·mL^-1 potassium dihydrogen phosphate acetonitrile（V∶V = 7∶3） as the mobile phase. The flow rate was 1.0 mL·min^-1，and the determination wavelength was 345 nm. The column temperature was 37 ℃，and the sample size was 20 μL. The HPLC was established for the determination of the 6 main components of Huang-Lian-Jie-Du decoction（genipin，berberine，coptisine，jateorhizine，baicalin and palmatine）. The standard curves of 6 components determined by HPLC were drawn，and the methodological evaluation（linear evaluation，recovery rate，accuracy and precision，stability） was carried out. The established HPLC was used to determine the samples of Huang-Lian-Jie-Du decoction. Results The linear ranges of genipin，berberine，coptisine，jateorhizine，baicalin and palmatine were 25.00-800.00，32.00-1 024.00，20.00-640.00，20.30-650.00，13.13-420.00 and 25.00-800.00 μg·mL^-1，respectively. The precision and accuracy were <2%. The recoveries were 105.30%，102.88%，103.51%，107.76%，96.29% and 99.34%，respectively. The stability test results showed that the peak area relative standard derivations（RSD） of genipin，berberine，coptisine，jateorhizine，baicalin and palmatine were all <2% after 24 h at room temperature，3 times frozen and thawing，2-8 ℃ for 1 week，and -80 ℃ for 1 month，and the determination results were all stable. The main components of Huang-Lian-Jie-Du decoction were determined by HPLC. The concentrations from high to low were jateorhizine（2 799.00 μg·mL^-1），berberine（2 358.03 μg·mL^-1），genipin（1 793.34 μg·mL^-1），palmatine（945.30 μg·mL^-1），coptisine （192.00 μg·mL^-1） and baicalin（171.00 μg·mL^-1）. Conclusions The established HPLC for the determination of the main components of Huang-Lian-Jie-Du decoction is rapid，simple，reproducible and accurate，which lays a foundation for the subsequent research and clinical application of Huang-Lian-Jie-Du decoction.

Key words: Huang-Lian-Jie-Du decoction, Jateorhizine, Berberine, Genipin, Palmatine, Coptisine, Baicalin, High performance liquid chromatography