Abstract: A method was established for the determination of D-chiro-inositol(D-CI)by pre-column derivatization HPLC. Repeatability,stability,precision and added standard recovery of the method and the derivatization conditions were investigated. Derivatization conditions were as follows:0.2 mL of benzoyl chloride,0.6 mL of pyridine,reaction temperature 70℃,reaction time of 60 min. Chromatographic conditions were as follows:using C18 column(4.6 mm×250 mm,5 μm),with acetonitrile and ultrapure water as mobile phrase in gradient elution,flow rate of 1.0 mL/min,column temperature of 30℃,and UV detection wavelength of 230 nm. The results showed that D-CI of buckwheat won good separation by the method,and the method got a good linear relationship of D-CI concentration and peak area at a range of 0.004~0.04 mg/mL with 0.999 of R² and the recovery was 98.40%~100.67%. So the method was stable,sebsitive,good reproducibility,and could be used to determine the content of D-CI from buckwheat.