Objective: To develop a UPLC-MS/MS method for the simultaneous determination of 18 constituents in Zishen Qinggan Daiping formula. Method: The samples were separated on a Acquity UPLC BEH C18 column (2.1 mm×100 mm

1.7 μm) by a gradient elution with the mobile phase of 0.1% formic acid-water and acetonitrileas at a flow rate of 0.3 mL · min-1. The column temperature was 50 ℃. The sample size was 5 μL. The electrospray ion source was adopted to monitor in the polarity in positive and negative mode. The multiple-reaction monitoring (MRM) was applied in the quantitative analysis. Result: Within 16 min

the 18 constituents (malic acid

citrate acid

gallic acid

oleanolicacid

ursolic acid

2

3

5

4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside

l-deoxynojirimycin

cassiaside

rubrofusarin gentiobioside

cassiaside B

resveratrol

physcion-8-O-β-D-glucopyranoside

aurantio-obtusin

chryso-obtusin

obtusinfolin

emodin

chrysophanol

icaritin) in Zishen Qinggan Daiping formula were completed separated. The regression equations showed a good linear relationship between the peak area and the concentration of each compound. The average recoveries ranged from 98.81% to 100.80%

and the precision ranged between 0.09% and 2.9%. Conclusion: The methodology validation shows that the method can be used for the content determination and quality control of 18 constituents in Zishen Qinggan Daiping formula.