Objective: To develop an HPLC method for determining the content of hesperidin and rosmarinic acid in Menthae Spicatae Herba. Method: The conditions of sample preparation

chromatography column and mobile phase were optimized. The linear ranges

reproducibility and recovery rate were evaluated based on a Thermo Scientific HPLC system to establish the method of quality control for determining the content of hesperidin and rosmarinic acid in Menthae Spicatae Herba. Result: HPLC analysis was conducted on a Inertsil ODS-3 C18 column (4.6 mm×250 mm

5 μm)

which was eluted with the mobile phase of acetonitrile-0.2% phosphoric acid (18: 82)

at a flow rate of 1.0 mL·min-1 under the temperature of 30 ℃

and the detection wavelength was set at 283 nm for hesperidin

and that of rosmarinic acid was under temperature of 35 ℃

and the detection wavelength was set at 330 nm. The regression equation of hesperidin was Y=27.61X+0.040 2 (r=0.999 9

0.008 434-1.687 μg) and that of rosmarinic acid was Y=51.44X-0.172 3 (r=0.999 9

0.018 91-1.891 μg);the average recovery of hesperidin was 96.43%(RSD 1.4%)

for rosmarinic acid it was 99.99%(RSD 1.7%). Conclusion: The method is simple

accurate

and repeatable

and so can be used for determining the content of hesperidin and rosmarinic acid in Menthae Spicatae Herba.