RP-HPLC simultaneous determination of three biological base in antler velvet

Objective:To establish an HPLC method for simultaneous and rapid determination of three main biological base components including uracil,hypoxanthine and uridine in antler velvet.Methods:An external standard method with Agilent TC C18 column(250 mm×4.6 mm,5 μm) was adopted with the mobile phase was consisted of 3% methanol-water solution containing 0.07% acetic acid at the flow rate of 1.0 mL·min-1;The detection wavelength was 254 nm and the column was 25 ℃.Results:Uracil,hypoxanthine and uridine were well separated by this method.Linearities of uracil,hypoxanthine and uridine were good(r=0.9995,0.9999,0.9996)in the range of 1.7-33.3 mg·L-1.The detection limits(S/N=3) were 9.375,18.75 and 75 μg·L-1 for uracil,hypoxanthine and uridine,respectively.The average recoveries of the three biological base components were found in the range of 98.4%-103.4%.Conclusion:The method is not only simple,rapid and presents good resolution and reproducibility,but also shows a high utility value,which has been successfully applied to the simultaneous determination of uracil,hypoxanthine and uridine in antler velvet.