Objective: The main active ingredients in Cephalotaxus hainanensis include cephalotaxine

harringtonine

homoharringtonine. This study aimed to establish a UPLC method for determination of cephalotaxine

harringtonine and homoharringtonine and analyze their metabolic accumulation and distribution in different tissues in C. hainanensis. Method: The method was performed on Agilent 1290Ⅱ UPLC platform. The suitable conditions of UPLC were Agilent Infinitylab EC-C18 column (2.1 mm×100 mm

2.7 μm

Agilent Infinitylab)

methanol and 20 mmol·L-1 ammonium carbonate aqueous solution as mobile phase

gradient elution at 0.8 mL·min-1

detective wavelength at 291 nm

injection volume of 5 μL

and column temperature at 30℃. Result: Under the above chromatographic conditions

the chromatogram baseline was stable

and each target compound had a good peak shape and resolution. The results showed a good linear relationship between content and peak area in all three C.hainanensis alkaloids. The methodological experiment showed that the instrument precision

method reproducibility and recovery rate all conform to the requirements of quantitative determination. Conclusion: This determination method has a good precision

stability and repeatability

with the advantages of shorter determination time

simpler operation and more accurate results. All of these three alkaloids could be detected in 12 min. It can be used to determine the contents of cephalotaxine

harringtonine and homoharringtonine in C. hainanensis

and establish a foundation for further study of metabolic accumulation in C. hainanensis.