柱前衍生反相高效液相色谱法测定马尾松(Pinus massoniana)针叶中γ-氨基丁酸和17种游离氨基酸

Determination of gamma-aminobutyric acid and seventeen free amino acids in Pinus massoniana Needles by reversed-phase high performance liquid chromatography with pre-column derivation

建立了自动在线柱前衍生反相高效液相色谱法同时测定γ-氨基丁酸（GABA）和17种游离氨基酸含量的方法。以邻苯二甲醛-9-芴基甲基氯甲酸酯（OPA—FMOC）为衍生试剂进行衍生，AgilentHypersilAA-ODS-C18色谱柱分离，梯度洗脱，二极管阵列检测器检测，在19min内分离测定了马尾松苗木针叶中GABA和17种游离氨基酸的含量。该方法测定氨基酸的回收率高于90．1％，精密度和重现性均较好（相对标准偏差为0．21％～2．81％），经测定，发现马尾松被马尾松毛虫取食后，所测18种氨基酸总量明显降低，从418．3μg·g^-1降低到310．4μg·g^-1鲜重。

An automated pre-column derivation procedure has been developed for the determination of gamma-aminobutyric acid （GABA） and 17 free amino acids by means of reversed-phase high performance liquid chromatography. GABA and the 17 free amino acids in needles of Pinus massoniana seedlings were determined with a 0-phthaldialdelhyde-9-fluore-nylmethyl chloroformate pre-column derivation in 19 min. The amino acid derivatives were separated on an Agilent Hypersil AA-ODS- C18 column by a gradient elution method and identified with a DAD detector. The recovery of amino acids by this method was more than 90. 1% with high precision and reproducibility （ its relative standard deviation ranged from 0.21% to 2. 81% ）. The total amount of amino acids in the needles of P. massoniana seedling decreased decidedly from 418.3 to 310.4μg·g-2 FW after insect damage.