邻苯二甲醛柱前衍生反相高效液相色谱法测定珍珠母中氨基酸

DETERMINATION OF AMINO ACIDS IN PEARL SHELL BY RP-HPLC WITH OPA PRE-COLUMN DERIVATIZATION

以邻苯二甲醛和 β-巯基乙醇为衍生化试剂 ,柱前衍生 ,ODS( C1 8)柱分离 ,p H5.6乙酸盐缓冲液、甲醇二元梯度洗脱 ,荧光检测 ,在 32 min内分离测定了珍珠母中 8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良好的线性关系 ,相关系数在 0 .9984～ 0 .9994之间。对于 8种氨基酸 ,标准加入回收率在 95%～ 1 0 5%之间。

A pre column derivatization procedure is described for the determination of amino acids in pearl shell by reversed phase high performance liquid chromatography. o Phthaldialdehyde/ β mercaptoethanol is used for the derivatization of amino acids. The separation of amino acid derivatives is accomplished on a column packed with 5μm ODS (octadecylsilance). A acetate buffer at pH 5.6 and 5% THF is used as the mobile phase A, while methyl alcohol is employed as the mobile phase B. Linear gradiemt elution is adopted with a fluorescent detector ( E x=340nm, E m=460nm). Eight amino acids in pearl shell are separated and detected in 32min. From the results, obtained a good linear relationship exists between peak area and concentrations of amino acids. The correlation coefficients are in the range of 0.9984~0.9994. The recoveries of 8 amino acids are in the range of 95%~105%.