A new type of agar microspheres with an average particle diameter of about 110 ?m was first prepared by water/oil emulsification of a polysaccharide agar solution.A yield about 88.6% can be achieved using this approach.Compared to commercially available agarose media,the agar media we prepared features much lower cost and enables a significant decrease of flow resistance due to the larger particle diameter,and is thus more suitable for large-scale applications.The agar microspheres were functionalized with polymeric β-cyclodextrin,and applied for isolation and purification of puerarin,a well-known traditional Chinese drug,from a crude extract of Radix puerariae (root of the plant Pueraria lobata).To the best of our knowledge,fewer studies on agar microspheres have been reported for application as chromatographic media.It was demonstrated that the polymeric β-cyclodextrin modified agar microspheres enable similar separation and selectivity to those previously obtained with 30 agarose gel media.The maximum dynamic loading capacity for puerarin is around 0.85 mg crude extract per mL packed gel.Puerarin with a purity of 96% could be recovered with a yield of 86%.