To find out the most reasonable analysis conditions of paeoniflorin, different paeoniflorin extraction methods and various UV detector wavelengths were conducted with paeonia radix of 4-year old Euisung local variety. The most reasonable paeoniflorin extraction time by reflux apparatus was 1hr. and by ultrasonic apparatus was 3hrs. and those methods were completed only once. Concentration of paeoniflorin by reflux apparatuses at 1hr. and 2hrs. of extracting time were higher than those of ultrasonic apparatus, and the differences were highly significant. However, the differences of paeoniflorin concentration at 3hrs. and 4hrs. in two methods were not significant. In comparing paeoniflorin concentration of many lines, ultrasonic extracting apparatus was more simple and effective than the reflux apparatus. Paeoniflorin was more reasonable sensitivity at 240nm, and albiflorin was 254nm by HPLC. When paeoniflorin and albiflorin were analyzed simultaneously, 254nm was more stable than any other wavelength.