Abstract: OBJECTIVE To screen out the most reliable and accurate method for Ch.P by comparison on three HPLC methods on the determination of cyclovirobuxium D and evaluation on the method's accuracy, intermediate precision and practicability. METHODS NP-HPLC method with SiO2 column and ELSD detection, NP-HPLC method with NH2 column and end-wavelength detection, and ion-pair RP-HPLC method with C18 column and end-wavelength detection were employed to determine cyclovirobuxium D in huangyangning tablets and raw materials, the method evaluation was given following the chromatographic features, content results, method sensitivities, accuracy and intermediate precision which were compared. RESULTS The raw materials separated by 3 methods could substantially occure 8, 10 and 14 peaks respectively, the recoveries (n=9) were 96.38% (RSD=2.6%), 75.24% (RSD=3.2%) and 97.97% (RSD=1.1%), and the intermediate precision of instruments expressed as RSD (n=3) were 3.9%, 7.1%, 4.0% for tablets and 3.5%, 4.3%, 4.7% for raw materials. CONCLUSION Among 3 methods, in terms of accuracy, intermediate precision and practicability, the ion-pair RP-HPLC method performed best. Therefore it could be adopted as a pharmacopoeia method.