Objective: This study was aimed to estabilish a high-performance liquid chromatography (HPLC) method for HPLC characteristics of aqueous extract of Zhenwu decoction (ZWT) and simultaneous determination of makers-characteristic chromatography

benzoylmesaconine

benzoylaconine

atractylenolide Ⅲ

paeoniflorin and 6-gingerol. Method: The separation was carried out on a Phenomenex Luna C18(4.6 mm×250 mm

5μm) column eluted with mobile phases of acetonitrile(A) and water containing 0.2% phosphoric acid(B) in gradient mode at the flow rate of 1.0 mL·min-1.The column temperature was maintained at 30℃ and the detection wavelength was set at 230 nm. Result: The obtained HPLC characteristics containing 36 common peaks

which were in well shape and high resolution. Five constituents of ZWT were analyzed and quantified paeoniflorin(1.410±0.024) g·L-1

benzoylmesaconine(0.066±0.001) g·L-1

benzoylaconine(0.015±0.001) g·L-1

atractylenolide Ⅲ(0.028±0.003) g·L-1

6-gingerol(0.069±0.001) g·L-1

with the linear range as follows Y=25.907X+9.741 (r=0.999 7)

Y=15.265X-3.202 (r=0.999 5)

Y=15.660X-4.699 (r=0.999 1)

Y=63.73X-10.740 (r=0.999 6)

Y=21.645X-4.074 (r=0.999 5)

respectively. The precision(1.72%

1.76%

2.78%

1.20%

1.05% for RSD of peak area

0.81%

0.47%

0.43%

0.07%

0.05% for RSD of retention time)

repeatability(1.95%

2.56%

2.93%

2.65%

3.01% for RSD of peak area;0.03%

0.19%

0.28%

0.11%

0.12% for RSD of retention time)

stability (1.05%

1.36%

2.89%

2.81%

2.64% for RSD of peak area;0.15%

0.12%

0.39%

0.04%

0.08% for RSD of retention time)

and average recovery (99.85%

101.67%

101.91%

99.82%

101.44%

respectively) of this method were all satisfied. Conclusion: The method established in this study could be used in simultaneous determination of the five makers of zhenwu decoction and could be used to control the quality of ZWT aqueous extract.