Determination of the Chlorogenic Acid in Callus and Suspension Cells of Eucommia ulmoides Oliv by HPLC

Objective To establish an HPLC method for the determination of chlorogenic acid in callus and suspension cells of Eucommia ulmoides Oliv.Methods The separation was performed on Hypersil C18-ODS(250mm×4.6mm,5μm) column,using acetonitrile-water-H3PO4(12∶87.9∶0.1)as mobile phase,detected at 327 nm.The flow rate was 1.0 ml/min,the column temperature was 25 ℃.Results The linear range of chlorogenic acid was 2.216～55.4 mg/ml,r=0.998 4,the average recovery of chlorogenic acid in casllus and suspension cells were 100.69% and 97.62%,RSD were 2.42% and 2.72%,respectively.Conclusion The method is rapid,simple,accurate and stable.Chlorogenic acid content in callus and suspension cells of Eucommia ulmoides Oliv are 1.528 mg/g and 3.949mg/g respectively.