The certified reference material (CRM) of honokiol was develpoed according to the technical requirements of CRM General and Statistical Principle for Certification (GB/T 15000.3-2008). Honokiol was separated and prepared by high-speed counter-current chromatography with crude extracts of Magnolia officinalis Rehd. et Wils. as material, and their structures were confirmed by UV, IR, MS, 1H and 13C NMR. Simultaneously, the purity and moisture content of Honokiol was analyzed by thin layer chromatography, LC-MS, and thermo-gravimetric to establish HPLC analysis technology. Honokiol was divided into bottles for homogeneity test, stability test, and cooperative certification by 8 laboratories. The results indicated that the homogeneity of honokiol samples were excellent, with good stability within 48 months at 0-4oC. The certified value of the reference material of honokiol was 99.72% with the expanded uncertainty of 0.22% in confidence coefficient of 95%. The reference material of honokiol was successfully developed, which reached the technical requirement of GB/T 15000.3-2008 and could be traceable. The CRM can be used in the content determination and testing method validation of honokiol related products.