色谱

色谱 ›› 2014, Vol. 32 ›› Issue (2): 184-188.DOI: 10.3724/SP.J.1123.2013.09027

• 技术与应用 • 上一篇    下一篇

高效液相色谱-电喷雾串联质谱法测定饲料中的N-氨基甲酰-L-谷氨酸

唐圣芸, 王远兴, 温平威, 辛贞   

  1. 南昌大学食品科学与技术国家重点实验室, 江西 南昌 330047
  • 收稿日期:2013-09-18 修回日期:2013-11-19 出版日期:2014-02-08 发布日期:2014-01-25
  • 通讯作者: 王远兴
  • 基金资助:

    南昌大学食品科学与技术国家重点实验室自由探索课题(SKLF-TS-200915).

Determination of N-carbamyl-L-glutamic acid in feedstuff by high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry

TANG Shengyun, WANG Yuanxing, WEN Pingwei, XIN Zhen   

  1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China
  • Received:2013-09-18 Revised:2013-11-19 Online:2014-02-08 Published:2014-01-25

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摘要：

建立了高效液相色谱-电喷雾串联质谱(HPLC-ESI-MS/MS)测定饲料中N-氨基甲酰-L-谷氨酸(NCG)含量的方法。饲料样品经甲醇提取、混合型强阴离子交换反相固相萃取(PXA)柱净化、HPLC分离后，采用ESI-MS/MS在正离子多反应监测(MRM)模式下进行检测，以碎片离子m/z 148.0和m/z 84.0进行定性，以碎片离子m/z 130.0进行定量。NCG的检出限(S/N >3)为24 μg/kg，定量限(S/N >10)为80 μg/kg，在20~1000 μg/L的质量浓度范围内峰面积与含量的线性关系良好，相关系数为0.9999。对饲料中NCG在80、200、500 mg/kg等3个添加水平下的回收率进行了测定，分别为104.0%、103.5%、95.3%，相对标准偏差分别为7.5%、6.3%、5.8%。结果表明，该方法操作简单，净化效果好，快速，灵敏度和准确度高，符合对饲料样品中NCG检测分析的要求。

关键词: N-氨基甲酰-L-谷氨酸, 高效液相色谱-电喷雾串联质谱法, 固相萃取, 饲料

Abstract:

A method was developed for the determination of N-carbamyl-L-glutamic acid (NCG) in feedstuff by high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The feedstuff samples were extracted with methanol, cleaned-up by a ProElut mixed-mode of strong anion exchange inverse column (PXA). Then the samples were separated with HPLC and detected with MS/MS in multiple reaction monitoring (MRM) mode via positive electrospray ionization (ESI+). The fragment ions of m/z 148.0 and m/z 84.0 were used for qualitative analysis, and m/z 130.0 was selected for quantitative analysis. The limit of detection for NCG was 24 μg/kg (S/N >3), and the limit of quantification was 80 μg/kg (S/N >10). The standard calibration curve was linear over the range of 20-1000 μg/L, and the correlation coefficient was 0.9999. The recoveries of NCG in feedstuff were 104.0%, 103.5%, 95.3%, at the three spiked levels of 80, 200, 500 mg/kg with the relative standard deviations (RSDs) of 7.5%, 6.3% and 5.8%. The results showed that the method is simple, fast, highly sensitive, specific and with good purification effect. It can be applied to the analysis and determination of NCG in feedstuff.

Key words: N-carbamyl-L-glutamic acid, feedstuff, high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS), solid phase extraction (SPE)

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