采用反相高效液相色谱法测定菜豆（豆科）种子中的β-氰基-L-丙氨酸、γ-谷氨酰基-β-氰基-L-丙氨酸和常见游离氨基酸。
Determination of β -Cyano-L-alanine, γ -Glutamyl- β -cyano-L-alanine, and Common Free Amino Acids in Vicia sativa (Fabaceae) Seeds by Reversed-Phase High-Performance Liquid Chromatography.
机构信息
Instituto de la Grasa (C.S.I.C.), Avenida Padre García Tejero 4, 41012 Sevilla, Spain.
出版信息
J Anal Methods Chem. 2014;2014:409089. doi: 10.1155/2014/409089. Epub 2014 Dec 22.
A method for determination of β-cyano-L-alanine, γ-glutamyl-β-cyano-L-alanine and other free amino acids in Vicia sativa is presented. Seed extracts were derivatized by reaction with diethyl ethoxymethylenemalonate and analyzed by reverse-phase high-performance liquid chromatography. Calibration curves showed very good linearity of the response. The limit of detection and quantification was 0.15 and 0.50 μM, respectively. The method has high intra- (RSD = 0.28-0.31%) and interrepeatability (RSD = 2.76-3.08%) and remarkable accuracy with a 99% recovery in spiked samples. The method is very easy to carry out and allows for ready analysis of large number of samples using very basic HPLC equipment because the derivatized samples are very stable and have very good chromatographic properties. The method has been applied to the determination of γ-glutamyl-β-cyano-L-alanine, β-cyano-L-alanine, and common free amino acids in eight wild populations of V. sativa from southwestern Spain.
本文介绍了一种用于测定 Vicia sativa 中β-氰基-L-丙氨酸、γ-谷氨酰基-β-氰基-L-丙氨酸和其他游离氨基酸的方法。种子提取物通过与二乙基乙氧基亚甲基丙二酸盐反应进行衍生化，并通过反相高效液相色谱法进行分析。校准曲线显示出非常好的响应线性。检测限和定量限分别为 0.15 和 0.50 μM。该方法具有较高的精密度（内重复性 RSD = 0.28-0.31%）和重现性（RSD = 2.76-3.08%），在加标样品中回收率达到 99%，准确度高。该方法非常简单，可使用非常基本的 HPLC 设备对大量样品进行快速分析，因为衍生化样品非常稳定，具有良好的色谱性能。该方法已应用于测定来自西班牙西南部的 8 个野生 V. sativa 种群中的 γ-谷氨酰基-β-氰基-L-丙氨酸、β-氰基-L-丙氨酸和常见游离氨基酸。
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