PITC柱前衍生高效液相色谱法测定接骨木果实中氨基酸含量

Determination of Amino Acids in the Fruits of Sambucus williamsii by PITC Pre-column Derivation and RP-HPLC Method

为了测定接骨木果实中游离氨基酸和水解氨基酸含量,采用PITC柱前衍生高效液相色谱法（RP-HPLC）以异硫氰酸苯酯（PITC）作柱前衍生试剂,采用RP-HPLC梯度洗脱的方法测定接骨木果实中的游离和水解氨基酸的峰面积并计算其含量。结果表明：接骨木果实中氨基酸和水解氨基酸的含量分别为2.107%和7.297%;18种氨基酸分离良好,线性关系良好,空白无干扰,精密度RSD在0.634%～2.532%,稳定性RSD在0.729%～2.774%,重复性RSD在0.664%～2.884%,平均回收率在95.49%～103.67%,且RSD均小于3%。RP-HPLC法灵敏、简便、快捷,能准确测定接骨木果实中游离氨基酸和水解氨基酸的含量。

For the determination of free and hydrolysis amino acids in the fruits of S. williarnsii, taking phenyl isothiocyanate （PITC） as the pre-column derivating reagent, RP-HPLC gradient elution method was employed to measure the peak area and contents of free and hydrolytic amino acid. Results： Contents of free amino acid and hydrolytic amino acid in the fruits of S. williarnsii were 2. 107% and 7. 297% respectively, separation of 18 kinds of amino acids was good, the linear relationship was good, and the blank had no interference, The RSD of precision was between 0. 634% and 2. 532%, the stability was between 0. 729Y0 and 2. 774%, the repeatability was between 0. 664% and 2. 884%, the sample recovery rate was between 95. 49% and 103. 67%, and the RSD was less than 3%. The method is sensitive, simple, fast and able to determine the content of elderberry and free amino acid hydrolysis accurately.