Determination of Angelica Sinensis Polysaccharide ( ASP1 ) in Rats by HPLC-FD with Pre-column Derivat-ization / 中国药师

ABSTRACT

Objective:To establish an HPLC-FD method for determining the content of Angelica sinensis polysaccharide (ASP1) to lay foundation for its pharmacokinetic study in rats. Methods: Purified ASP1 was labeled with FITC by the method of Belder and Granath to obtain ASP1-FITC. The tissue samples were treated with 30% trichloroacetic acid and 11% NaOH before injection. The samples were determined by HPLC-FD. A PL aquagel-OH MIXED column was used,and the mobile phase was phosphate buffer( dis-solve NaH2PO32. 34g , Na2HPO34. 33 g and NaCl 11. 70 g into 1000 ml water) with pH of 7. 0. The flow rate was 0. 5 ml·min-1. The excitation and emission wavelengths was set at 495 nm and 520 nm, respectively. Results:The linear calibration curve was within the concentration range of 0. 25-40. 00 μg·ml-1(r=0. 9996)with the lower limit of quantification of 0. 20μg·ml-1 in tissue sam-ples. The extraction recovery of ASP1 was determined at low, medium and high concentration with the recovery of 91. 98%-114. 20%. The intra and inter-day RSDs were lower than 8. 31% and 2. 94%, respectively. Conclusion:The method to determine the content of ASP1 in rats by HPLC-FD with pre-column derivatization has been esablished. It is simple, accurate and reliable, which can be suc-cessfully applied in the study of pharmacokinetics and tissue distribution of ASP1 in rats.