邻苯二甲醛柱前衍生反相高效液相色谱法测定水牛角中氨基酸的含量

Determination of Amino Acids in Buffalo Horn by Reversed-Phase High Performance Liquid Chromatography with o-Phthaldialdelhyde Pre-colomn Derivatization

以邻苯二甲醛和β-巯基乙醇为衍生化试剂，往前衍生，ODS（C18）柱分离；pH5．6醋酸盐缓冲溶液、甲醇二元梯度洗脱，荧光检测，在32min内分离测定了水牛角中12种氨基酸的含量。在一定浓度范围内，氨基酸的峰面积与其浓度呈现良好的线性关系，相关系数在0．9975－0．9994之间。对于12种氨基酸，标准加入回收率在94％－106％之间。

A pre-column derivatization procedure is descrided for the determination of amino acids in Buffalo horn by reversed-phase high performance liquid chromatography, o-phthaldialdelhyde/β-mercaptoethano was used for the derivatizaion of amino acids. The separation of amino acid derivatives was accomplished on a column packed with 5 μm ODS(octadecylsilance). An acetate buffer at pH 5. 6was used as the mobile phase A, while methanol was employed as the mobile phase B. Linear gradient elution was adopted with a fluonaent detector(Ex = 340 nm, Em = 460 nm). Twelve amino acids in buffalo hom were separated and detected in 32 min. From the results, a good linear relationship exists between peak area and concentration of amino acids ranged from 0 ~ 0. 1 g/L to 0 ~ 0. 5 g/L. The correlation coefficients are in the range of 0. 9975 ~ 0. 9994. The recovery of 12 amino acids is in the range of 94% ~ 100%.