柱前衍生反相高效液相色谱法测定小麦中氨基酸含量

Simultaneous Determination of Amino Acids in Wheat by Reversed Phase High Performance Liguid Chromatography with Precolumn Derivatization

以肌氨酸为内标物,邻苯二甲醛-9-芴甲基氯甲酸酯为柱前衍生剂,用ODS色谱柱在柱温40℃下,采用二元梯度洗脱,DAD检测器在338 nm波长处检测,建立了一种利用反相高效液相色谱同时测定小麦籽粒中17种氨基酸的方法.氨基酸浓度在5～800 μmol/L范围内,其峰面积与内标物峰面积的比值和氨基酸浓度的线性相关系数均大于0.996;17种氨基酸的加标回收率在97.5%～103.1%范围内.应用本方法对小麦籽粒中氨基酸含量进行测定,取得了较理想的结果.同时,本法还可应用于糙米和玉米等粮食中氨基酸含量的测定.

A reversed-phase high performance liquid chromatographic method for simultaneous determination of 17 amino acids was investigated. The sample was mixed with sarcosine as the internal standard, then derivatized with o-phthaldialdehyde-9-fluorenylmethyl chloroformate and analyzed on a ODS column at 40℃ using gradient elution with diode array detection at 338 nm. When the amino acid concentration was at 5 - 800 μmol/L, range, the correlation coetiicients between the ratios of peak area of amino acid to that of the internal standard, and the amino acid concentrations were above 0.996. The recoveries of standard addition of amino acids were 97.5% - 103.1%. This method was applied to the separation of 17 amino acids in wheat kernel, and determiantion of amino acids in grains such as brown rice and maize with satisfactory results.