Simultaneous Determination of Four Amino Acids in Fufang Ejiao Buxue Granule by HPLC

[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a Waters Sunfire C 18 column(4.6 mm×250 mm,5μm).A binary mobile solvent was used:mobile solvent A was acetonitrile-0.1 mol/L sodium acetate solution(adjusting pH to 6.5 with 36%acetic acid)(7∶93)and mobile solvent B was acetonitrile-H 2O(4∶1).The mobile phase was delivered at a flow rate of 1.0 mL/min with a gradient elution profile(0-13 min,100%A→93%A;13-17.9 min,93%A→88%A;17.9-29.0 min,88%A→85%A;29-39 min,85%A→66%A;39-45 min,66%A→0%A).The column temperature was at 43℃.The detection wavelength was 254 nm.[Results]The injection volume of L-hydroxyproline,glycine,alanine,and L-proline showed a good linear relationship with the chromatographic peak area in the range of 0.012 to 0.117,0.022 to 0.218,0.010 to 0.097,0.016 to 0.160μg,separately.The average recovery rate(n=6)was 96.4%,97.3%,97.1%,and 99.4%,respectively;the relative standard deviations were 1.2%,1.9%,1.7%,and 0.9%,respectively.[Conclusions]This method is simple in operation and good in reproducibility,and provides a reliable method for controlling the quality of Fufang Ejiao Buxue Granules.