Simultaneous determination of hydroxysafflor yellow A and ferulic acid in rat plasma after oral administration of the co‐extractum of Rhizoma chuanxiong and Flos Carthami by HPLC–diode array detector

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A simple, rapid and accurate HPLC method has been developed for simultaneous determination of hydroxysafflor yellow A and ferulic acid in rat plasma after oral administration of the co-extractum of Rhizoma chuanxiong and Flos Carthami. Plasma samples were deproteinized with 6% perchloric acid, and riboflavin was used as internal standard. The supernatant after centrifuge was injected into a Shimadzu C(18) (150 x 4.6 mm, i.d. 5 microm) column. Gradient elution for A:B (0 min, 90:10; 25 min, 70:30; 27 min, stop) was applied. The mobile phase was composed of 0.022 mol/L potassium dihydrogen phosphate solutions, adjusted to pH 3.0 with phosphoric acid for pump A, and 90% (v/v) acetonitrile for pump B. The assay was shown to be linear over the range 0.046-4.6 microg/mL (r(2) = 0.9995) for hydroxysafflor yellow A and 0.037-3.7 microg/mL (r(2) = 0.9998) for ferulic acid. Mean recovery was 97.5% for hydroxysafflor yellow A and 83.6% for ferulic acid. Both of the intra-day and inter-day precisions were <or=8.1%. The limits of quantification of hydroxysafflor yellow A and ferulic acid were 0.046 and 0.037 microg/mL, respectively. The HPLC method was successfully applied to the pharmacokinetic studies of hydroxysafflor yellow A and ferulic acid in rat plasma after oral administration of the co-extractum of Rhizoma chuanxiong and Flos Carthami.