高效液相色谱法测定发芽麦粒中γ-氨基丁酸(GABA)含量

Determination of γ-Aminobutyric Acid(GABA) in Germinated Wheat Seed by HPLC

建立了一种测定发芽麦粒中γ-氨基丁酸(GABA)含量的HPLC方法,并对四川10个小麦品种中GABA含量差异及其发芽培养期间变化规律进行了分析。采用FDNB(2,4-二硝基氟苯)为柱前衍生剂,色谱条件为:色谱柱Luna C18_((2))(150 mm×4.60 mm,5μm),乙腈-0.02 mmol/L乙酸铵混合溶液为流动相(V:V=15:85),柱温30℃,流速1.0 mL/min,测定波长320 nm。结果表明,在0.025 5~0.127 6μg范围内,呈良好的线性关系,最低检出限1.081×10^(-5)μg/mL,方法的平均加标回收率为97%。采用建立的方法测定不同小麦品种在萌发12 h后的GABA含量在0.238~5.26 mg/100 g之间,不同品种间GABA的含量差异显著,在萌发的0~12 h内,GABA含量呈现先上升后下降的趋势,在8 h时含量达到最高。

The HPLC determination method for γ- aminobutyric acid （GABA） in germinated wheat seed has been improved in the paper. Meanwhile, the GABA content in 10 different kinds of wheat cultivars from Sichuan area have been investigated by HPLC method. Isocratic elution was used for separation and quantification of GABA deri- rates with FDNB. GABA was determined on Luna C18（2） （ 150 mm × 4.6 mm, 5 μm） with mobile phase of acetate ammonium （0.02 mmol/L） and acetonitrile （V： V = 85：15）. The flow rate was 1.0 mL/min; column temperature 30 ℃, and the detection wave length, was 320 nm respectively. The results indicated that a good linearity was ob- served within a range from 0. 025 5 -0. 127 6 μg. The limit of detection is 1. 081 × 10^-5 μg/mL. The average recov- eries are 97% for GABA determination. GABA content ranged 0. 238 - 5.26 rag/100 g in different wheat varieties af- ter germination 12 h was determined by the established method. The significant difference of content in GABA among different varieties was also observed. During germination of 0 - 12 h, GABA content has shown a trend of first in- creased and then decreased, with content peak at 8 h.