SPE-HPLC-MS/MS测定水稻中氨基酸

Determination of Amino Acids in Rice by High Performance Liquid Chromatography Tandem Mass Spectrometry

建立同时测定水稻中异亮氨酸、蛋氨酸、酪氨酸等10种氨基酸的固相萃取-高效液相色谱-串联质谱方(SPE-HPLC-MS/MS)法,为水稻中多种氨基酸含量检测提供技术参考。水稻经液氮研磨并高速冷冻离心后,样品提取液经过固相萃取小柱净化和富集,用乙腈-水溶液进行复溶。以Zic-HILIC(150 mm×2.1 mm,3.5μm)为色谱柱,以乙腈和0.1%甲酸水溶液为流动相进行梯度洗脱。质谱采用电喷雾离子源电离,以正离子扫描,多反应监测模式检测。结果显示,10种氨基酸在2.00×10^(-3)~2.00μg·kg^(-1)范围内线性良好,相关系数(R2)均大于0.9980,检出限在1.00×10-4~6.40×10-4μg·kg^(-1)之间;在0.01,0.20和2.00μg kg^(-1)加标水平下,平均回收率在82.0%~118.2%之间,相对标准偏差(SRSD)在3.2%~8.7%之间(n=6)。该方法快速、灵敏,适合同时分析水稻中异亮氨酸、蛋氨酸、酪氨酸、脯氨酸、谷氨酰胺、谷氨酸、天冬氨酸、精氨酸、赖氨酸和鸟氨酸10种氨基酸。

A solid phase extraction high performance liquid chromatography tandem mass spectrometry(SPE-HPLC-MS/MS)method for the simultaneous determination of amino acid in rice was established,which provided effective method to detect the content of amino acid.After the samples were ground by liquid nitrogen and centrifuged at high speed,they were separated by Zic-HILIC(150 mm×2.1 mm,3.5μm),and acetonitrile and 0.1%formic acid solution were used as mobile phases for isocratic elution.Positive electrospray ionization with multiple reactions monitoring(MRM)mode was used for detection.The results showed that the 10 kinds of amino acids had good linearity within the mass concentration range from 2.00×10^(-3)μg·kg^(-1)to 2.00μg·kg^(-1),the correlation coefficients(R2)were all greater than 0.9980,and the limits of detected(LOD)were from 1.00×10-4μg·kg^(-1)to 6.40×10-4μg·kg^(-1);The recoveries of the spiked levels(0.01,0.20 and 2.00μg·kg^(-1))were from 82.0%to 118.2%,and the relative standard deviation(SRSD)between 3.2%and 8.7%(n=6).The method was fast,sensitive and accurate,and was suitable for isoleucine,methionine,tyrosine,proline,glutamine,glutamate,asparagine,arginine,lysine and ornithine in rice tissue simultaneous analysis.