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Sensitive Analysis of 33 Free Amino Acids in Serum, Milk, and Muscle by Ultra-High Performance Liquid Chromatography-Quadrupole-Orbitrap High Resolution Mass Spectrometry

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Abstract

Quantitative analysis of 33 free amino acids (AAs) in serum, milk, and muscle samples was achieved using ultra-high performance liquid chromatography-quadrupole-orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). The samples were extracted using methanol and derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate to enhance chromatographic retention. Thirty-three AAs were separated on a 1.7-μm BEH C18 column within 15 min, which is at least six times faster than that of an AA analyzer. Serum, milk, and muscle samples spiked with free AAs were tested in terms of linearity, sensitivity, repeatability, and recovery. Results indicated that recoveries for the AAs ranged from 77.7 to 124.8 % with relative standard deviations (RSD) less than 12.3 %. The lower limit of detection ranged from 0.003 to 0.68 μmol/L and 0.003 to 0.27 nmol/g for body fluid and muscle samples, respectively. Finally, the developed method was used for the determination of free AAs in actual serum, milk, and muscle samples with good results.

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Acknowledgments

The animal samples were kindly obtained from the Institute of Subtropical Agriculture of the Chinese Academy of Sciences (Changsha, China). The financial support from the National Basic Research Program of China (973 Program, 2013CB127305), the Special Fund for Agro Scientific Research in the Public Interest (201203088), and the National Natural Science Foundation of China (31472126) are gratefully acknowledged.

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    Authors and Affiliations

    1. State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing, 100193, People’s Republic of China

      Bingjie Yin, Tingting Li & Pingli He

    2. Beijing Huiwen Middle School, Beijing, 100061, People’s Republic of China

      Siran Zhang

    3. State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, People’s Republic of China

      Zhen Li

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    1. Bingjie Yin
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    2. Tingting Li
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    Correspondence to Pingli He.

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    All the authors declare that they have no conflict of interest in the present work.

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    In this study, all applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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    Bingjie Yin and Tingting Li contributed equally to this work.

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    Yin, B., Li, T., Zhang, S. et al. Sensitive Analysis of 33 Free Amino Acids in Serum, Milk, and Muscle by Ultra-High Performance Liquid Chromatography-Quadrupole-Orbitrap High Resolution Mass Spectrometry. Food Anal. Methods 9, 2814–2823 (2016). https://doi.org/10.1007/s12161-016-0463-0

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