柱前衍生RP-HPLC法测定犀牛角中氨基酸的含量

目的]建立柱前衍生法测定犀牛角中氨基酸含量的方法。方法]犀牛角以酸水解法制备水解氨基酸样品溶液,采用柱前衍生-液相色谱法(RP-HPLC)法测定氨基酸含量。色谱条件为:色谱柱为Waters Nova-Pak C18柱(150 mm×3.9 mm,5μm),以乙腈-缓冲盐为流动相进行梯度洗脱,流速为1.0 ml/min,紫外检测器检测波长为248 nm,柱温为37℃。结果]17种氨基酸在0.003 4～0.191mg/ml范围内有良好的线性关系(r0.999 4),平均回收率(n=6)为97.8%。结论]犀牛角中含有大量的氨基酸,其总氨基酸含量高达41.3%。该方法对研究犀牛角的药用价值与寻找犀牛角药用替代物有积极意义。

Determination of Amino Acids in Rhinoceros Horn by Pre-column Derivatization RP-HPLC

Objective] To determine the contents of amino acids in rhinoceros horn by pre-column derivation.Method] Acid hydrolysis method was used for the extraction of hydrolytic amino acid in rhinoceros horn.Then using pre-column derivation RP-HPLC,the contents of amino acids in rhinoceros horm were determined.The chromatographic conditions:Waters Nova-Pak C18column(150 mm×3.9 mm,5 μm)was used,the acetonitrile-buffered saline as mobile phase was used for gradient elution,with a flow rate of 1.0 ml/min,and the wavelength was 248 nm,the column temperature was 37 ℃.Result] The calibration curve was linear in the range of 0.003 4~0.191 mg/ml of amino acids(r>0.999 4).The average recovery of amino acids was 97.8%.Conclusion] Rhinoceros horn contains abundant amino acids with a total amount of 41.3%.So the method is helpful to the study of medicinal value and finding an alternative of rhinoceros horn.