ABSTRACT
Objective To establish an HPLC-DAD method for simultaneous determination of main anti-oxidant constituents in leaves and stem of Rosmarinus officinalis. Methods The separation was performed on an Agilent Zorbax SB-Aq C18 column(250 mm × 4.6 mm,5 μm), using methanol (A) and potassium dihydrogen phosphate solution (B) (pH 3.0) as the mobile phase at the flow rate of 1.0 mL/min for a gradient elution. The detection wavelength was set at 328 nm for the detection of chlorogenic acid, caffeic acid, rosmarinic acid, luteolin, apigenin, and genkwanin, and 284 nm for the detection of paeonol, rosmanol, hesperetin, carnosol, and carnosic acid. Column temperature was set at 30 ℃, and the volume of sample injection was 10 μL. Results The linear range was 2.02-64.64 μm/mL for chlorogenic acid, 1.74-55.68 μmol/mL for caffeic acid, 2.76-88.32 μmol/mL for rosmarinic acid, 0.24-7.68 μmol/mL for paeonol, 0.46-14.72 μmol/mL for rosmanol, 0.22-7.04 μmol/mL for hesperetin, 0.72-23.04 μmol/mL for luteolin, 3.74-119.68 μmol/mL for apigenin, 5.1-163.28 μmol/mL for carnosol, 0.4-12.8 μmol/mL for genkwanin, and 5.22-167.04 μmol/mL for carnosic acid. The average recoveries of the 11 constituents were from 98.1%-100.5%, and the RSD values were 0.67%-2.14%. Conclusion The method is simple and accurate,which can be used for quality and quantity analysis of the main anti-oxidant constituents in leaves and stem of R. officinalis.