Simultaneous determination of danshensu,protocatechuic acid and protocatechuic aldehyde in rabbit plasma by HPLC

Aim To establish an high performance liquid chromatography(HPLC) method for simultaneous determination of danshensu,protocatechuic acid and protocatechuic aldehyde in rabbit plasma.Methods Trichloroacetic acid was used to remove the protein in plasma,and the resulted supernatant was extracted by acetic ether.Then,danshensu,protocatechuic acid and protocatechuic aldehyde were determined by reverse HPLC.Results The detection limits of danshensu,protocatechuic acid and protocatechuic aldehyde were 0.03、0.08 and 0.04 μg·mL-1,respectively,and the linear range were 0.06～40.0 μg·mL-1,0.16～100.0 μg·mL-1 and 0.08～50.0 μg·mL-1,respectively.The recovery of them in low,middle and high concentration was in the range of 82.7%～102.0%,and the precision of inter-and intra-day was among the range of 0.56%～4.84%.Conclusion This method could be used for simultaneous determining danshensu,protocatechuic acid and protocatechuic aldehyde in plasma,with the properties of high sensitivity and specificity.Thus,this method could be used for pharmacokinetic study on danshensu,protocatechuic acid and protocatechuic aldehyde.